WO2000046349A1 - Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells - Google Patents
Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells Download PDFInfo
- Publication number
- WO2000046349A1 WO2000046349A1 PCT/US2000/002688 US0002688W WO0046349A1 WO 2000046349 A1 WO2000046349 A1 WO 2000046349A1 US 0002688 W US0002688 W US 0002688W WO 0046349 A1 WO0046349 A1 WO 0046349A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- cells
- stromal
- matrix
- hemopoietic stem
- stem cells
- Prior art date
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0634—Cells from the blood or the immune system
- C12N5/0647—Haematopoietic stem cells; Uncommitted or multipotent progenitors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/14—Scaffolds; Matrices
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
- C12N11/089—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
- C12N11/096—Polyesters; Polyamides
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K2035/124—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells the cells being hematopoietic, bone marrow derived or blood cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/125—Stem cell factor [SCF], c-kit ligand [KL]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/145—Thrombopoietin [TPO]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/20—Cytokines; Chemokines
- C12N2501/26—Flt-3 ligand (CD135L, flk-2 ligand)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2502/00—Coculture with; Conditioned medium produced by
- C12N2502/13—Coculture with; Conditioned medium produced by connective tissue cells; generic mesenchyme cells, e.g. so-called "embryonic fibroblasts"
- C12N2502/1394—Bone marrow stromal cells; whole marrow
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/30—Synthetic polymers
Definitions
- the present invention relates to a method and apparatus for maintenance and expansion of hemopoietic stem cells. More particularly, the present invention relates to a three dimensional stromal cell plug flow bioreactor for the maintenance and/or expansion of hemopoietic stem cells and/or for the production of a conditioned medium for the maintenance and/or expansion of hemopoietic stem cells.
- the hemopoietic system in mammals is composed of a heterogenous population of cells that range in function from mature cells with limited proliferative potential to pluripotent stem cells with extensive proliferative, differentiative and self renewal capacities (1-3).
- Hemopoietic stem cells (HSC) are exclusively required for hemopoietic reconstitution following transplantation and serve as a primary target for gene therapy.
- stem cells are intimately associated in vivo with discrete niches within the marrow (4-6), which provide molecular signals that collectively mediate their differentiation and self renewal, via cell-cell contacts or short-range interactions (7).
- HIM hemopoietic inductive microenvironment
- Marrow stromal cells e.g., macrophages, fibroblasts, adipocytes and endothelial cells (8).
- Marrow stromal cells maintain the functional integrity of the HIM by providing extracellular matrix (ECM) proteins and basement membrane components that facilitate cell-cell contact (9-11). They also provide various soluble or resident cytokines needed for controlled hemopoietic cell differentiation and proliferation (12-14).
- ECM extracellular matrix
- a recently developed human stem cell assay detects a SCID repopulating cell (SRC), which homes to the bone marrow of non-obese diabetic (NOD)/SCID mice (27), where it gives rise to human myeloid, lymphoid, erythroid and CD34+ progenitor populations (28-30).
- SRC SCID repopulating cell
- NOD non-obese diabetic
- SCID mice non-obese diabetic mice
- the SRC is exclusively found in hemopoietic cell fractions expressing the CD34+38- surface antigen (31) and its frequency in CB (1/3x10 ⁇ cells) is enriched as compared to BM ( 1/9x10$ cells) or mobilized PB (1/ 6x10 ⁇ cells) (32).
- Very recent studies showed that the SRC resides within a subpopulation of CD34+/38-/CXCR4+ cells (33).
- CXCR4 a surface receptor for the chemokine stromal cell-derived factor 1 (SDF-1, 34), is apparently essential for homing and engraftment of human hemopoietic stem cells in the NOD/SCID marrow (33).
- SDF-1 chemokine stromal cell-derived factor 1
- allogeneic human marrow stroma was found to induce short-term (7-day) SRC maintenance, followed by a rapid, marked decline (6-fold) in activity (26).
- the inability to support the long-term maintenance/expansion of transplantable human stem cells on stromal cell layers may be attributed to several factors related to in vitro cultures of these cells. Among these, one may include the use of stromal cell monolayers, which do not reflect the in vivo growth conditions within the natural, three-dimensional structure of the bone marrow. Such conditions may diminish the capacity of stromal cells to provide the optimal, appropriate supportive microenvironment, as well as the capacity of stem cells to localize in specific niches and to physically interact with stromal cells and their products.
- a plug flow bioreactor system which closely mimics the 3D bone marrow microenvironment and which is capable of supporting the growth and prolonged maintenance of stromal cells, has been developed.
- the latter were seeded on porrosive inorganic carriers made of a non woven fabric matrix of polyester (54), enabling the propagation of large cell numbers in a relatively small volume.
- the structure and packing of the carrier have a major impact on oxygen and nutrient transfer, as well as on local concentrations and released stromal cell, products (e.g., ECM proteins, cytokines, 55).
- the capacity of stromal cells cultured in this system to promote the maintenance/expansion of transplantable human hemopoietic stem cells via direct cell-cell contact has been determined to be far superior over prior art methods.
- the capacity of conditioned medium of stromal cells cultured in this system to promote the maintenance/expansion of transplantable human hemopoietic stem cells via novel stromal-cell associated stem cell factors included therein has been determined to be far superior over prior art methods.
- a method of expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells comprising the steps of (a) obtaining undifferentiated hemopoietic stem cells or progenitor cells; and (b) seeding the undifferentiated hemopoietic stem cells or progenitor cells into a stationary phase plug-flow bioreactor in which a three dimensional stromal cell culture has been pre-established on a substrate in the form of a sheet, the substrate including a non-woven fibrous matrix forming a physiologically acceptable three-dimensional network of fibers, thereby expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells.
- the method further comprising the step of isolating the undifferentiated hemopoietic stem cells or progenitor cells.
- a method of expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells comprising the steps of (a) obtaining undifferentiated hemopoietic stem cells or progenitor cells; and (b) culturing the undifferentiated hemopoietic stem cells or progenitor cells in a medium containing a stromal cell conditioned medium, the stromal cell conditioned medium being derived from a stationary phase plug-flow bioreactor in which a three dimensional stromal cell culture has been established on a substrate in the form of a sheet, the substrate including a non-woven fibrous matrix forming a physiologically acceptable three-dimensional network of fibers, thereby expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells.
- a method of preparing a stromal cell conditioned medium useful in expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells comprising the steps of (a) establishing a stromal cell culture in a stationary phase plug-flow bioreactor on a substrate in the form of a sheet, the substrate including a non-woven fibrous matrix forming a physiologically acceptable three-dimensional network of fibers, thereby expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells; and (b) when a desired stromal cell density has been achieved, collecting medium from the stationary phase plug-flow bioreactor, thereby obtaining the stromal cell conditioned medium useful in expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells.
- a method of transplanting undifferentiated hemopoietic stem cells or progenitor cells into a recipient comprising the steps of (a) expanding/maintaining the undifferentiated hemopoietic stem cells or progenitor cells by (i) obtaining undifferentiated hemopoietic stem cells or progenitor cells; and (ii) seeding the undifferentiated hemopoietic stem cells or progenitor cells into a stationary phase plug-flow bioreactor in which a three dimensional stromal cell culture has been pre- established on a substrate in the form of a sheet, the substrate including a non-woven fibrous matrix forming a physiologically acceptable three- dimensional network of fibers, thereby expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells; and (b) transplanting the undifferentiated hemopoietic stem cells or progenitor cells resulting from step (a) in the recipient.
- the method further comprising the step of isolating the undifferentiated hemopoietic stem cells or progenitor cells prior to step (b).
- a method of transplanting undifferentiated hemopoietic stem cells or progenitor cells into a recipient comprising the steps of (a) expanding/maintaining the undifferentiated hemopoietic stem cells or progenitor cells by (i) obtaining undifferentiated hemopoietic stem cells or progenitor cells; and (ii) culturing the undifferentiated hemopoietic stem cells or progenitor cells in a medium containing a stromal cell conditioned medium, the stromal cell conditioned medium being derived from a stationary phase plug- flow bioreactor in which a three dimensional stromal cell culture has been established on a substrate in the form of a sheet, the substrate including a non- woven fibrous matrix forming a physiologically acceptable three-dimensional network of fibers, thereby expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells.
- a bioreactor plug comprising a container having an outlet and an inlet and containing therein a substrate in the form of a sheet, the substrate including a non-woven fibrous matrix forming a physiologically acceptable three-dimensional network of fibers, the substrate supporting at least 5 x 10 ⁇ stromal cells per cubic centimeter of the substrate.
- a plug-flow bioreactor comprising the above bioreactor plug.
- the undifferentiated hemopoietic stem cells or progenitor cells are cells isolated from a tissue selected from the group consisting of cord blood, mobilized peripheral blood and bone-marrow.
- the undifferentiated hemopoietic stem cells or progenitor cells and stromal cells of the stromal cell culture share common HLA antigens.
- the undifferentiated hemopoietic stem cells or progenitor cells and stromal cells of the stromal cell culture are from a single individual.
- the undifferentiated hemopoietic stem cells or progenitor cells and stromal cells of the stromal cell culture are from different individuals. According to still further features in the described preferred embodiments the undifferentiated hemopoietic stem cells or progenitor cells and stromal cells of the stromal cell culture are from the same species.
- the undifferentiated hemopoietic stem cells or progenitor cells and stromal cells of the stromal cell culture are from different species.
- stromal cells of the stromal cell culture are grown to a density of at least 5 x 10 ⁇ cells per a cubic centimeter of the substrate.
- stromal cells of the stromal cell culture are grown to a density of at least 10 ⁇ cells per a cubic centimeter of the substrate.
- the step of seeding the undifferentiated hemopoietic stem cells or progenitor cells into the stationary phase plug-flow bioreactor is effected while flow in the bioreactor is shut off for at least 10 hours following the seeding.
- the fibers form a pore volume as a percentage of total volume of from 40 to 95 % and a pore size of from 10 microns to 100 microns.
- the matrix is made of fiber selected from the group consisting of flat, non-round, and hollow fibers and mixtures thereof, the fibers being of from 0.5 microns to 50 microns in diameter or width.
- the matrix is composed of ribbon formed fibers having a width of from 2 microns
- the ratio of width to thickness of the fibers is at least 2: 1.
- the matrix having a pore volume as a percentage of total volume of from 60 to 95%.
- the matrix has a height of 50-1000 ⁇ m.
- the material of the matrix is selected from the group consisting of polyesters, polyalkylenes, polyfluorochloroethylenes, polyvinyl chloride, polystyrene, polysulfones, cellulose acetate, glass fibers, and inert metal fibers.
- the matrix is in a shape selected from the group consisting of squares, rings, discs, and cruciforms.
- the matrix is coated with poly-D-lysine.
- the present invention successfully addresses the shortcomings of the presently known configurations by providing more effective means for expanding/maintaining undifferentiated hemopoietic stem cells.
- Implementation of the method and bioreactor of the present invention may involve performing or completing selected tasks or steps manually, automatically, or a combination thereof.
- several selected steps could be implemented by hardware or by software on any operating system of any firmware or a combination thereof.
- selected steps of the invention could be implemented as a chip or a circuit.
- selected steps of the invention could be implemented as a plurality of software instructions being executed by a computer using any suitable operating system.
- selected steps of the method and bioreactor of the invention could be described as being performed by a data processor, such as a computing platform for executing a plurality of instructions.
- FIG. 1 is a schematic depiction of an exemplary plug-flow bioreactor 20 which served while reducing the present invention to practice; 1- medium reservoir; 2 - gas mixture container; 3 - gas filters; 4 - injection points; 5 - plug or container of plug flow bioreactor 20; 6 - flow monitors; 6a - flow valves; 7 - conditioned medium collecting/separating container; 8 - container for medium exchange; 9 - peristaltic pump; 10 - sampling point; 11- container for medium exchange; 12 - O2 monitor; 14 - steering device; PH - pH probe.
- FIG. 2 demonstrates CAFC maintenance by 14F1J cells.
- Cord blood CD34+ cells were seeded at limiting dilution onto irradiated 14F1J or primary human marrow stroma. Cobblestone formation was determined 5 weeks later. Results represent the mean ⁇ SD of 2 independent experiments.
- FIG. 3 demonstrates LTC-IC maintenance by 14F1J cells.
- Cord blood CD34+ cells were seeded at limiting dilution onto irradiated 14F1 J or primary human marrow stroma. Myeloid colony formation was determined 7 weeks later.
- FLT-3 ligand 300 ng/ml
- TPO 300 ng/ml
- SCF 100 ng/ml
- FIGs. 5a-b show scanning electron micrographs (SEM) of carrier seeded with 14F1 J stromal cell line following 10 days ( Figure 5a) or following 40 days ( Figure b). Magnification: x 150.
- FIGs. 6a-b demonstrate the effect of 3D versus 2D 14F1 J conditioned medium on CD34+38- expansion.
- CD34+ cells were seeded in suspension cultures in the presence of various concentrations of conditioned medium from 14F1 J and primary human marrow stroma.
- CD34+38- cell numbers were determined by FACS analysis. Results represent the mean ⁇ SD of 2 independent experiments.
- FIG. 7 demonstrates maintenance of CD34+38- cells on stromal- cell coated carriers.
- Stromal cell-coated carriers were removed from the 3D system into silicone-coated 96-well dishes, followed by addition of 1.5 x l ⁇ 4 CD34+ cells.
- Controls contained carriers alone and carrier- equivalent numbers of monolayer (2D) grown 14F1J cells. Cells were harvested at the designated times and analyzed by FACS. Results represent the mean ⁇ SD of 2 independent experiments.
- the present invention is of methods and bioreactor for hemopoietic stem cell expansion/maintenance which can be used for transplantation in a recipient or for other purposes as if further detailed hereinunder.
- the present invention is of a three dimensional stromal cell plug flow bioreactor for the maintenance and/or expansion of hemopoietic stem cells and/or for the production of a conditioned medium for the maintenance and/or expansion of hemopoietic stem cells, which can be used in a variety of applications.
- a novel three dimensional (3D) plug flow bioreactor which closely mimics the bone marrow microenvironment and which is capable of supporting the growth and prolonged maintenance of marrow stromal cells is described herein.
- the latter are seeded on porrosive carriers made of a non woven fabric matrix of polyester, packed in a glass column, thereby enabling the propagation of large cell numbers in a relatively small volume.
- the bioreactor was seeded with the murine 14F1 J stromal cell line or alternatively with primary human marrow stromal cells.
- the carriers contained a 100-fold increased cell density.
- the density at various levels of the column was the same, indicating a homogenous transfer of oxygen and nutrients to the cells.
- Media conditioned by stromal cells within the bioreactor (3D SCM) was superior to stromal cell monolayer (2D) SCM, in supporting the long-term maintenance of human cord blood (CB) CD34+38- cells.
- 3D SCM was also capable of supporting the expansion of CD34+38-CXCR4+ cells, which represent SCID/NOD repopulating cells (SRC).
- 3D SCM In the presence of cytokines (FLT3 ligand and TPO), 3D SCM enhanced stem cell self renewal and inhibited differentiation, while the opposite effect was induced by 2D SCM + cytokines.
- Three dimensional stromal-stem cell cocultures also exhibited superior maintenance of CD34+38- cells than cocultures on monolayer stromal cells.
- the human HSC is an essential target for transplantation and gene therapy.
- the bioreactor described herein is unique in that it combines both 3D stromal cell cultures with a continuous flow system. While 3D stromal-hemopoietic cell systems without continuous medium flow have recently been described (U.S. Pat. No. 5,541,107), the findings described herein (see, for example, Figure 7) clearly demonstrate the diminished advantage of 3D stromal cell cultures relative to monolayers, in the absence of continuous flow.
- the 3D plug-flow bioreactor described herein is capable of supporting the long-term growth of stromal cell lines, as well as primary marrow stromal cells.
- the use of stromal cells in the bioreactor is not only essential for the establishment of superior stromal-stem cell contact (via unique "niches" and cell-cell, cell-ECM interactions), but also for stromal cell production of known and novel soluble and membrane- bound cytokines.
- Stromal cells can facilitate the supplementation of such bioreactors with appropriate cytokines, by using genetically engineered cytokine-producing variants.
- Bioreactor stromal cells can also be engineered to serve as retroviral packaging cell lines, enabling the efficient transduction of genetic material into stem cells, within the bioreactor itself.
- the use of various stromal cells in the bioreactor can also allow the selection of the most suitable substrate for purging of Ph-positive stem cells, the latter known for their lesser capacity for stromal cell adherence (63).
- Primary stromal cells have the advantage that they enable the establishment of "autologous" stromal-stem cell bioreactors, on which autologous or even cord blood stem cells can be expanded and which eliminate the need to remove stromal cells prior to transplantation.
- the bioreactor of the present invention employs a growth matrix that substantially increases the available attachment surface for the adherence of the stromal cells so as to mimic the mechanical infrastructure of bone marrow. For example, for a growth matrix of 0.5 mm in height, the increase is by a factor of at least from 5 to 30 times, calculated by projection onto a base of the growth matrix.
- the matrix is used in sheet form, preferably non- woven fiber sheets, or sheets of open-pore foamed polymers
- the preferred thickness of the sheet is about 50 to 1000 ⁇ m or more, there being provided adequate porosity for cell entrance, entrance of nutrients and for removal of waste products from the sheet.
- the pores having an effective diameter of 10 ⁇ m to 100 ⁇ m.
- Such sheets can be prepared from fibers of various thicknesses, the preferred fiber thickness or fiber diameter range being from about 0.5 ⁇ m to 20 ⁇ m, still more preferred fibers are in the range of 10 ⁇ m to 15 ⁇ m in diameter.
- the structures of the invention may be supported by, or even better bonded to, a porous support sheet or screen providing for dimensional stability and physical strength.
- Such matrix sheets may also be cut, punched, or shredded to provide particles with projected area of the order of about 0.2 mm ⁇ to about 10 mm2, with the same order of thickness (about 50 to 1000 ⁇ m).
- the present invention provides expanded undifferentiated hemopoietic stem cell population which can be used in a variety of applications, such as, but not limited to: (i) expansion of human stem cells (of autologous or cord blood source) on recipient stroma, without the need for stromal-stem cell separation prior to transplantation; (ii) depletion of Ph+ CML stem cells in an autologous setting via stromal-stem cell interactions; (iii) gene transfer into self- renewing stem cells within the bioreactor or following harvesting from the bioreactor; (iv) production of 3D stromal cell conditioned medium (SCM) for ex-vivo maintenance/expansion of undifferentiated hemopoietic stem cells in suspension cultures or in a stem cell bioreactor; (v) isolation of novel proteins inducing stem cell self renewal in the absence of differentiation, as well as proteins having additional biological functions; (vi) isolation of 3D stromal cell RNA for cloning of novel stromal cell-
- SCM 3D stromal cell
- a method of expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells is effected by implementing the following method steps. First, undifferentiated hemopoietic stem cells or progenitor cells are obtained.
- the undifferentiated hemopoietic stem cells or progenitor cells are seeded into a stationary phase plug-flow bioreactor, an example of which is depicted in Figure 1 along with reference numerals, in which a three dimensional stromal cell culture, of either stromal cell line or primary stromal cell culture, have been pre- established on a substrate in the form of a sheet, the substrate including a non-woven fibrous matrix forming a physiologically acceptable three- dimensional network of fibers, thereby, as is further described above and exemplified in the Examples section that follows, expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells.
- undifferentiated hemopoietic stem cells refers to uncommitted hemopoietic cells.
- progenitor cells refers to committed, yet immature hemopoietic cells.
- Both undifferentiated hemopoietic stem cells and progenitor cells are CD34+ cells.
- the phrase "obtaining undifferentiated hemopoietic stem cells or progenitor cells” and its equivalent phrase “undifferentiated hemopoietic stem cells or progenitor cells are obtained” both refer to the obtainment of either isolated undifferentiated hemopoietic stem cells and/or progenitor cells, or a population of CD34+ cells which contain undifferentiated hemopoietic stem cells and progenitor cells.
- expanding and “expansion” refer to substantially differentiationless cell growth, i.e., increase of a cell population without differentiation accompanying such increase.
- the terms “maintaining” and “maintenance” refer to substantially differentiationless cell renewal, i.e., substantially stationary cell population without differentiation accompanying such stationarity.
- the term “differentiation” refers to change from relatively generalized to specialized kinds during development. Cell differentiation of various cell lineages is a well documented process and requires no further description herein.
- ex-vivo refers to cells removed from a living organism and are propagated outside the organism (e.g., in a test tube).
- the now expanded undifferentiated hemopoietic stem cells or progenitor cells can be isolated by a variety of affinity separation/labeling techniques, such as, but not limited to, fluorescence activated cell sorting and affinity separation via an affinity substrate.
- affinity separation/labeling techniques such as, but not limited to, fluorescence activated cell sorting and affinity separation via an affinity substrate.
- Affinity molecules which can be used to implement such isolation methods include anti-CD34 antibodies, for example, which bind CD34+ cells.
- the method according to this aspect of the present invention is effected by implementing the following method steps. First, undifferentiated hemopoietic stem cells or progenitor cells are obtained.
- the undifferentiated hemopoietic stem cells or progenitor cells are cultured in a medium containing, as a sole ingredient or as an additive, a stromal cell conditioned medium, the stromal cell conditioned medium being derived from a stationary phase plug- flow bioreactor in which a three dimensional stromal cell culture, of either stromal cell line or primary stromal cell culture, have been established on a substrate in the form of a sheet, the substrate including a non-woven fibrous matrix forming a physiologically acceptable three- dimensional network of fibers, thereby, as is further described above and exemplified in the Examples section that follows, expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells.
- a method of preparing a stromal cell conditioned medium useful in expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells is effected by implementing the following method steps.
- a stromal cell culture of either stromal cell line or primary stromal cell culture, is established in a stationary phase plug-flow bioreactor on a substrate in the form of a sheet, the substrate including a non-woven fibrous matrix forming a physiologically acceptable three-dimensional network of fibers, thereby expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells, second, when a desired stromal cell density has been achieved, say, for example, above 5 x 10 ⁇ or above 10 ⁇ cells per cubic centimeter of the matrix, collecting medium from the stationary phase plug-flow bioreactor, as is further described above and exemplified in the Examples section that follows, obtaining the stromal cell conditioned medium useful in expanding/maintaining undifferentiated hemopoietic stem cells or progenitor cells.
- a method of transplanting undifferentiated hemopoietic stem cells or progenitor cells into a recipient is effected by implementing the following method steps.
- Second, the undifferentiated hemopoietic stem cells or progenitor cells resulting from the first step are transplanted in the recipient.
- a bioreactor plug comprising a container 5, typically in the form of a column, having an outlet and an inlet and containing therein a substrate in the form of a sheet, the substrate including a non-woven fibrous matrix forming a physiologically acceptable three-dimensional network of fibers, the substrate supporting at least 5 x 10 ⁇ stromal cells, preferably, at least 10?, of either stromal cell line or primary stromal cell culture, per cubic centimeter of the substrate.
- a plug-flow bioreactor comprising the above bioreactor plug.
- the substrate may theoretically support up to 5 x 10 ⁇ cells per cubic centimeter thereof. Once sufficient cells have accumulated on the substrate, means such as irradiation can be employed to cease further cell growth, so as to control the exact number of cells supported by the substrate.
- the undifferentiated hemopoietic stem cells or progenitor cells which are used as a source for such cells while implementing the methods of the present invention can be purified or isolated from a tissue, such as, but not limited to, cord blood, cytokine-mobilized peripheral blood (collected by, for example, leukapheresis) and bone-marrow, all of which are known to include undifferentiated hemopoietic stem cells or progenitor cells.
- a tissue such as, but not limited to, cord blood, cytokine-mobilized peripheral blood (collected by, for example, leukapheresis) and bone-marrow, all of which are known to include undifferentiated hemopoietic stem cells or progenitor cells.
- Methods of such separation are well known in the art, the most frequently used being fluorescence activated cell sorting in which cells are first tagged by affinity labeling with a fluorophore and are than collected.
- the undifferentiated hemopoietic stem cells or progenitor cells and stromal cells of the stromal cell culture share common HLA antigens.
- the undifferentiated hemopoietic stem cells or progenitor cells and the stromal cells of the stromal cell culture are from a single individual. Thus, separation of cells is not required in case of transplantation thereof to a recipient.
- the undifferentiated hemopoietic stem cells or progenitor cells and stromal cells of the stromal cell culture are from different individuals.
- a future recipient of the undifferentiated hemopoietic stem cells or progenitor cells and stromal cells be used to provide the stromal cells, whereas the undifferentiated hemopoietic stem cells or progenitor cells and stromal cells are from a donor selected according to HLA compatibility to donate such cells to the recipient.
- HLA compatibility to donate such cells to the recipient.
- the undifferentiated hemopoietic stem cells or progenitor cells and stromal cells of the stromal cell culture are from the same species.
- the undifferentiated hemopoietic stem cells or progenitor cells and stromal cells of the stromal cell culture are from different species.
- the step of seeding the undifferentiated hemopoietic stem cells or progenitor cells into the stationary phase plug-flow bioreactor is effected while flow in the bioreactor is shut off for at least 10 hours following such seeding, so as to enable the cells to anchor to the stromal cell covered matrix.
- the fibers of the substrate form a pore volume as a percentage of total volume of from 40 to 95 % and a pore size of from 10 microns to 100 microns.
- the matrix making the substrate is made of fiber selected from the group consisting of flat, non-round, and hollow fibers and mixtures thereof, the fibers being of from 0.5 microns to 50 microns in diameter or width.
- the matrix is composed of ribbon formed fibers having a width of from 2 microns.
- the ratio of width to thickness of the fibers is at least 2:1.
- the matrix making the substrate having a pore volume as a percentage of total volume of from 60 to 95 %.
- the matrix has a height of 50-1000 ⁇ m, whereas stacks thereof are employed.
- the material of the matrix making the substrate is selected from the group consisting of polyesters, polyalkylenes, polyfluorochloroethylenes, polyvinyl chloride, polystyrene, polysulfones, cellulose acetate, glass fibers, and inert metal fibers.
- the matrix is in a shape selected from the group consisting of squares, rings, discs, and cruciforms.
- the matrix is coated with poly-D-lysine.
- Bioreactor The bioreactor used in accordance with the teachings of the present invention was constructed in accordance with the design described in Figure 1.
- the glassware was designed and manufactured at the Technion (Israel) and connected by silicone tubing (Degania, Israel).
- PBS phosphate buffered saline
- the PBS was removed via container [8] and the bioreactor was circulated in a 37°C incubator with 300 ml Dulbecco's high-glucose medium (DMEM; GIBCO BRL) containing 10 % heat-inactivated fetal calf serum (FCS; Beit Ha'Emek Industries, Israel) and a Pen-Strep-Nystatin mixture (100 U/ml: 100 ⁇ g/ml:1.25 ⁇ n/ml; Beit Ha'Emek), for a period of 48 hours. Circulating medium was replaced with fresh DMEM containing the above + 2 mM L-glutamine (Beit Ha'Emek).
- DMEM Dulbecco's high-glucose medium
- FCS heat-inactivated fetal calf serum
- Pen-Strep-Nystatin mixture 100 U/ml: 100 ⁇ g/ml:1.25 ⁇ n/ml; Beit Ha'Emek
- Stromal cells Stromal cell lines were maintained at 37°C in DMEM supplemented with 10 % FCS, in a fully humidified incubator of 5 % CO2 in air. Cells were grown in tissue culture flasks (Corning) and were split by trypsinization upon reaching confluence. Primary human marrow stromal cultures were established from aspirated sternal marrow of hematologically healthy donors undergoing open-heart surgery. Briefly, marrow aspirates were diluted 3-fold in Hank's Balanced Salts Solution (HBSS; GIBCO BRL) and were subject to Ficoll-Hypaque (Robbins Scientific Corp. Sunnyvale, CA) density gradient centrifugation.
- HBSS Hank's Balanced Salts Solution
- Marrow mononuclear cells ( ⁇ 1.077 gm/cm ⁇ ) were collected, washed 3 times in HBSS and resuspended in long-term culture (LTC) medium, consisting of DMEM supplemented with 12.5 % FCS, 12.5 % horse serum (Beit Ha'Emek), 10 M ⁇ -mercaptoethanol (Merck) and 10-6 mol/L hydrocortwasone sodium succinate (Sigma).
- LTC long-term culture
- Cells were incubated in 25 ml tissue culture flasks (Corning) for 3 days at 37 °C (5 % CO2) and then at 33 °C (idem) with weekly culture refeeding. Stromal cells from individual donors were employed for each bioreactor.
- stromal cells were split by trypsinization (0.25 % Trypsin and EDTA in Puck's Saline A; Beit Ha'Emek) every 10 days, to allow sufficient stromal cell expansion.
- trypsinization 0.25 % Trypsin and EDTA in Puck's Saline A; Beit Ha'Emek
- stromal cells were irradiated (1500 cGy) using a 137c s source, cultures were maintained at 33 °C in LTC medium.
- stromal cells Confluent cultures of stromal cell lines or 5 -week primary marrow stromal cells were trypsinized and the cells washed 3 times in HBSS, resuspended in bioreactor medium (see above), counted and seeded at 10 ⁇ cells/ml in 10 ml volumes via an injection point ([4], Figure 1) onto 10 ml carriers in the glass column of the bioreactor. Immediately following seeding, circulation was stopped for 16 hours to allow the cells to settle on the carriers. Stromal cell growth in the bioreactor was monitored by removal of carriers and cell enumeration by the MTT method (56). When stromal cells were confluent, medium was replaced with LTC medium, for continued studies (preparation of SCM, stem cell seeding).
- stromal cell conditioned medium At equivalent cell densities, monolayer and bioreactor stromal cells were recharged with fresh LTC culture medium. SCM was collected following overnight incubation of the cells. For this purpose, medium flow in the 3D cultures was stopped for 16 hours and removed directly from the column prior to re-initiation of circulation. For analysis of the effect of CD34+ cells on stromal cell production of SRC, circulation was stopped at various intervals (2-7 days) after seeding of CD34+ into the 3D system and medium collected from the column as described above. SCM was spun (1000 x g, 10 minutes), filtered and stored at -20 °C. Stromal cells were also grown in the bioreactor in serum-free medium, for the collection of SCM, thereby excluding undefined variables.
- SCM stromal cell conditioned medium
- CD34+ cells Umbilical cord blood samples taken under sterile conditions during delivery were fractionated on Ficoll- Hypaque and buoyant ( ⁇ 1.077 gr/cm ⁇ ) mononuclear cells collected. Cells from individual CB samples were pooled, incubated with anti- CD34 antibodies and isolated by midi MACS (Miltenyl Biotech). Suspension cultures of CD34+ cells: CB CD34+ cells (5xlONwell) were incubated in 24-well dishes (TPP, Switzerland), in 0.5 ml of 0-100 % SCM, minus or plus 300 ng/ml each of FLT3 ligand, SCF, or TPO, alone or combined. Controls contained LTC medium plus or minus cytokines.
- Cells were incubated at 37 °C at 5 % CO2 in air. Culture medium was exchanged weekly. Prior to seeding and at various times (1-3 weeks), cells were harvested, enumerated and assayed for CD34+/38-/CXCR4+ by flow cytometry. Output assays can also include SRC, CAFC and LTC-IC. Stromal-stem cell cocultures: Isolated, pooled CB CD34+ cells were seeded at equivalent numbers (about 5 x 10 ⁇ ) onto monolayer or bioreactor containing equivalent densities of confluent stromal cells.
- Output assays can also include SRC, CAFC and LTC-IC.
- LTC-IC and CAFC assays Freshly isolated CD34+ cells, cells isolated from stromal-stem cell cocultures or from suspension cultures, were assayed for LTC-IC and CAFC, as previously described (16, 17). Confluent primary marrow stromal cells were trypsinized, irradiated (1500 cGy) and plated in 0J ml in 96-well dishes (Corning) at 1.5xl ⁇ 4/well. 24 replicate wells/group were established. Stromal cells were overlaid with 0J ml of LTC medium containing serial dilutions of CD34+ cells (500-5 cells/well), or with serial dilutions of cells harvested from various assays.
- the percentage of wells with at least one phase-dark hemopoietic clone of at least five cells (cobblestone area) beneath the stromal layer was determined at week 6 following seeding of the test cell suspensions, in serial dilutions.
- the bioreactor system employed while reducing the present invention to practice is depicted in Figure 1. It contained four parallel plug flow bioreactor units [5]. Each bioreactor unit contained 1 gram of porrosive carriers (4 mm in diameter) made of a non woven fabric matrix of polyester (58). These carriers enable the propagation of large cell numbers in a relatively small volume. The structure and packing of the carrier have a major impact on oxygen and nutrient transfer, as well as on local concentrations and released stromal cell products (e.g., ECM proteins, cytokines, 59). The bioreactor was maintained in an incubator of 37 °C.
- each bioreactor contains a sampling and injection point [4], allowing the sequential seeding of stromal and hemopoietic cells.
- Culture medium was supplied at pH 7.0 [13] from a reservoir [1].
- the reservoir was supplied by a filtered [3] gas mixture containing air/C02/02 [2] at differing proportions in order to maintain 5-40 % dissolved oxygen at exit from the column, depending on cell density in the bioreactor.
- the O2 proportion was suited to the level of dissolved O2 at the bioreactor exit, as was determined by a monitor [12].
- the gas mixture was supplied to the reservoir via silicone tubes.
- the culture medium was passed through a separating container [7] which enabled collection of circulating, nonadherent cells. Circulation of the medium was obtained by means of a peristaltic pump [9] operating at a rate of 0J-3 ml/minute.
- the bioreactor units were equipped with an additional sampling point [10] and two containers [8, 1 1] for continuous medium exchange at a rate of 10-50 ml/day.
- the use of four parallel bioreactor units enables periodic dismantling for purposes such as cell removal, scanning electron microscopy, histology, immunohistochemistry, RNA extraction, etc.
- the carriers When seeded into the bioreactor at 1.5x10 ⁇ cells/10 ml culture volume, 14F1 J cells grew and spread on the carriers (Figure 5). By day 40 following seeding, the carriers contained a 100-fold increased cell density, i.e., approximately 1.5x10 ⁇ cells/carrier, 1.5x10? cells/ml (Table 1).
- MTT analysis included 5 carriers/determination. Mean of 2 independent experiments. The cellular density on carriers at various levels of the column was the same, indicating a homogenous transfer of oxygen and nutrients to the cells.
- the culture conditions were optimized for these cells: culture medium (Dulbecco's high-glucose medium + 10 % fetal calf serum), flow rate (1 ml/min), medium exchange frequency (once a week), initial seeding density (as above). No beneficial effect was found for collagen or poly L-lysine carrier coating, on the growth rate and final density of 14F1 J cells.
- Preliminary findings with primary human marrow stromal cells (Table 1) indicated a similar density of 14F1 J and primary stromal cells, on days 10 and 14 following seeding, respectively.
- CD34+38- cells in suspension cultures seeded with human CB CD34+ cells was determined.
- the activity was compared to SCM obtained from monolayer cultures (2D SCM) containing the same concentration of stromal cells.
- SCM from 14F1J cells was found to be equally or more capable of supporting the maintenance of human CB CD34+38- cells, than SCM from primary marrow stromal cells.
- a maximal effect of 14F1J SCM was consistently observed at a lower concentration than that of primary marrow SCM.
- 3D SCM was found to be superior to 2D SCM of both cell types, in supporting the expansion of human CB CD34+38- cells. The difference in activity between 2D and 3D SCM was more pronounced with culture duration (14 versus 21 days).
- Human CB CD34+ cells (8 x 10 /point) were seeded in suspension cultures containing LTC medium or 50% 3D 14F1 J SCM. Cultures were harvested 7 days later and cells analyzed by FACS. CD34+38- and CD34+38-CXCR4+ inputs were 2800 and 1 12, respectively.
- Table 3 demonstrates the effect of cytokines in suspension cultures of CD34+ containing 2D versus 3D SCM. The results clearly demonstrate that 3D SCM was superior to 2D SCM in supporting the maintenance of both CD34+38- and more importantly, the CD34+38- CXCR4+ (SRC) subset.
- Human CB CD34+ cells (2.6xl0 /point) 50% 2D vs 3D 14F1.1 SCM, in the absence or presence of FLT3 ligand (300 ng/ml) TPO (300 ng/ml) or SCF (50 ng/ml). Cultures were harvested 7 days later and cells analyzed by FACS. CD34+38- and CD34+38-CXCR4+ inputs were 7900 and 360, respectively.
- TPO+FLT3 ligand reduced the yield of CD34+38-/ CD34+38-CXCR4+ in the presence of 2D SCM but enhanced their yield in cultures supplemented with 3D SCM. Again, this can be attributed to the lesser extent of differentiation in the 3D system, as determined by the CD34+ surface marker.
- SCF induced a marked increase in stem cell differentiation and a marked decline in the yield of CD34+38-/ CD34+38-CXCR4+ cells.
- CD34+38- cells were seeded into a bioreactor containing 4 columns of non-irradiated, 14Fl J-coated carriers, in 350 ml circulating culture medium. Medium flow was stopped for 16 hours and continued thereafter at a normal rate (1 ml/min). Following 4 days of coculture, circulating medium contained 10 % of the initially seeded CD34+38- cells, determined by FACS analysis of harvested viable cells. Following 18 days of culture, circulating medium contained 0.4 % CD34+38- cells, while carrier -adherent cells contained 3 % of the initially seeded CD34+38- population.
- Varnum-Finney B Purton LE, Yu M, Brashem-Stein C, Flowers D, Pope S, Moore KA, Le Roux I, Mann R, Gray G, Artavanis- Tsakonas S, Bernstein ID. Blood 91 :4084, 1998.
Abstract
Description
Claims
Priority Applications (19)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020017009869A KR20020013496A (en) | 1999-02-04 | 2000-02-04 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and(or) progenitor cells |
NZ513303A NZ513303A (en) | 1999-02-04 | 2000-02-04 | Three dimensional plug flow bioreactor, and its use for the maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
US09/890,401 US6911201B1 (en) | 1999-02-04 | 2000-02-04 | Method of producing undifferentiated hemopoietic stem cells using a stationary phase plug-flow bioreactor |
DE60043534T DE60043534D1 (en) | 1999-02-04 | 2000-02-04 | METHOD AND DEVICE FOR STABILIZING AND EXPANSIONING HEMATOPOIETIC STEM CELLS AND / OR PRE-CELL CELLS |
EP00913340A EP1147176B1 (en) | 1999-02-04 | 2000-02-04 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
CA2360664A CA2360664C (en) | 1999-02-04 | 2000-02-04 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
JP2000597409A JP4523169B2 (en) | 1999-02-04 | 2000-02-04 | Method and apparatus for maintaining and increasing hematopoietic stem cells and / or progenitor cells |
BR0009403-0A BR0009403A (en) | 1999-02-04 | 2000-02-04 | Method of expansion / conservation of undifferentiated hemopoietic stem cells or progenitor cells, method of preparing a conditioned stomach cell medium useful in the expansion / conservation of undifferentiated hemopoietic stem cells or progenitor cells, method of transplanting undifferentiated hemopoietic stem cells or progenitor cells into a container , buffer of bioreactor and bioreactor |
AT00913340T ATE452181T1 (en) | 1999-02-04 | 2000-02-04 | METHOD AND DEVICE FOR MAINTAINING AND EXPANSING HEMATOPOIETIC STEM CELLS AND/OR PROCURSOR CELLS |
MXPA01007820A MXPA01007820A (en) | 1999-02-04 | 2000-02-04 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells. |
AU34807/00A AU759719B2 (en) | 1999-02-04 | 2000-02-04 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
IL144629A IL144629A (en) | 1999-02-04 | 2001-07-30 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
HK02107728.2A HK1046154B (en) | 1999-02-04 | 2002-10-24 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
US11/102,654 US20050180958A1 (en) | 1999-02-04 | 2005-04-11 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
US11/102,635 US20050176137A1 (en) | 1999-02-04 | 2005-04-11 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
US11/102,623 US7678573B2 (en) | 1999-02-04 | 2005-04-11 | Method of preparing a conditioned medium from a confluent stromal cell culture |
US11/102,625 US7534609B2 (en) | 1999-02-04 | 2005-04-11 | Method of expanding undifferentiated hemopoietic stem cells |
US12/230,566 US20090004738A1 (en) | 1999-02-04 | 2008-09-02 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
US13/360,068 US20120122220A1 (en) | 1999-02-04 | 2012-01-27 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US11878999P | 1999-02-04 | 1999-02-04 | |
US60/118,789 | 1999-02-04 |
Related Child Applications (6)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US09890401 A-371-Of-International | 2000-02-04 | ||
EP09174118.1A Previously-Filed-Application EP2208782B1 (en) | 1999-02-04 | 2000-02-04 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
US11/102,623 Continuation-In-Part US7678573B2 (en) | 1999-02-04 | 2005-04-11 | Method of preparing a conditioned medium from a confluent stromal cell culture |
US11/102,625 Continuation-In-Part US7534609B2 (en) | 1999-02-04 | 2005-04-11 | Method of expanding undifferentiated hemopoietic stem cells |
US11/102,654 Continuation-In-Part US20050180958A1 (en) | 1999-02-04 | 2005-04-11 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
US11/102,635 Continuation-In-Part US20050176137A1 (en) | 1999-02-04 | 2005-04-11 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2000046349A1 true WO2000046349A1 (en) | 2000-08-10 |
Family
ID=22380750
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/US2000/002688 WO2000046349A1 (en) | 1999-02-04 | 2000-02-04 | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
Country Status (18)
Country | Link |
---|---|
US (7) | US6911201B1 (en) |
EP (3) | EP1147176B1 (en) |
JP (1) | JP4523169B2 (en) |
KR (1) | KR20020013496A (en) |
CN (1) | CN100402642C (en) |
AT (1) | ATE452181T1 (en) |
AU (1) | AU759719B2 (en) |
BR (1) | BR0009403A (en) |
CA (1) | CA2360664C (en) |
DE (1) | DE60043534D1 (en) |
ES (1) | ES2338405T3 (en) |
HK (1) | HK1046154B (en) |
IL (1) | IL144629A (en) |
MX (1) | MXPA01007820A (en) |
NZ (1) | NZ513303A (en) |
RU (1) | RU2249039C2 (en) |
WO (1) | WO2000046349A1 (en) |
ZA (1) | ZA200106483B (en) |
Cited By (21)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2000069449A2 (en) * | 1999-05-14 | 2000-11-23 | Advanced Tissue Sciences, Inc. | Conditioned cell culture medium compositions and methods of use |
GB2366295A (en) * | 2000-05-10 | 2002-03-06 | Tristem Ireland Ltd | Device and methods to prepare undifferentiated cells |
WO2002024874A2 (en) * | 2000-09-21 | 2002-03-28 | Schering Corporation | Methods for preparing interferon producing dentitric cells |
WO2003104386A1 (en) * | 2002-05-22 | 2003-12-18 | 株式会社エムビーエス | Culture apparatus, artificial tissue and blood preparation |
US7160726B2 (en) | 2001-06-07 | 2007-01-09 | Skin Medica, Inc. | Compositions comprising conditioned cell culture media and uses thereof |
EP2010647A2 (en) * | 2006-03-23 | 2009-01-07 | Pluristem Ltd. | Methods for cell expansion and uses of cells and conditioned media produced thereby for therapy |
EP2208782A3 (en) * | 1999-02-04 | 2010-11-03 | Pluristem Ltd. | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
US8492140B2 (en) | 2002-04-08 | 2013-07-23 | Octane Biotech Inc. | Automated tissue engineering system |
US8846393B2 (en) | 2005-11-29 | 2014-09-30 | Gamida-Cell Ltd. | Methods of improving stem cell homing and engraftment |
US9175266B2 (en) | 2012-07-23 | 2015-11-03 | Gamida Cell Ltd. | Enhancement of natural killer (NK) cell proliferation and activity |
US9567569B2 (en) | 2012-07-23 | 2017-02-14 | Gamida Cell Ltd. | Methods of culturing and expanding mesenchymal stem cells |
US9758762B2 (en) | 2008-09-02 | 2017-09-12 | Pluristem Ltd. | Perfusion bioreactor for culturing CD200—placenta adherent cells |
US10047345B2 (en) | 2012-02-13 | 2018-08-14 | Gamida-Cell Ltd. | Culturing of mesenchymal stem cells with FGF4 and nicotinamide |
EP3406272A1 (en) | 2015-01-26 | 2018-11-28 | Ube Industries, Ltd. | Cell culture method using bone marrow-like structure, and porous polymide film for healing bone injury site |
EP3464565A4 (en) * | 2016-05-25 | 2020-01-01 | Terumo BCT, Inc. | Cell expansion |
WO2021110908A1 (en) | 2019-12-04 | 2021-06-10 | Centre Hospitalier Universitaire Vaudois (C.H.U.V.) | Device and process for tissue-engineering and regenerative medicine |
US11371018B2 (en) | 2017-09-01 | 2022-06-28 | Octane Biotech Inc. | End-to-end cell therapy automation |
US11597905B2 (en) | 2018-12-28 | 2023-03-07 | Octane Biotech Inc. | Cell culture and tissue engineering systems with controlled environmental zones |
US11714096B2 (en) | 2018-12-21 | 2023-08-01 | Octane Biotech Inc. | Carousel for modular biologic production units |
US11718833B2 (en) | 2018-12-21 | 2023-08-08 | Lonza Walkersville, Inc. | Automated production of viral vectors |
US11773365B2 (en) | 2019-02-08 | 2023-10-03 | Lonza Walkersville, Inc. | Cell concentration methods and devices for use in automated bioreactors |
Families Citing this family (79)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040042997A1 (en) * | 2002-09-03 | 2004-03-04 | Donnie Rudd | Method of regenerating human tissue |
WO2004090121A2 (en) * | 2003-04-08 | 2004-10-21 | Yeda Research And Development Co. Ltd | Stem cells having increased sensitivity to a chemoattractant and methods of generating and using same |
US8352031B2 (en) | 2004-03-10 | 2013-01-08 | Impulse Dynamics Nv | Protein activity modification |
US7622108B2 (en) * | 2004-04-23 | 2009-11-24 | Bioe, Inc. | Multi-lineage progenitor cells |
CN101080486B (en) * | 2004-04-23 | 2012-05-16 | 佰欧益股份有限公司 | Multi-lineage progenitor cells |
US7521221B2 (en) * | 2005-11-21 | 2009-04-21 | Board Of Trustees Of The University Of Arknasas | Staphylococcus aureus strain CYL1892 |
ES2549111T3 (en) | 2005-12-29 | 2015-10-23 | Anthrogenesis Corporation | Placental stem cell populations |
EP1994143A4 (en) * | 2006-02-06 | 2009-08-26 | Pluristem Ltd | Method and apparatus for maintenance and expansion of hematopoietic stem cells from mononuclear cells |
CN100384987C (en) * | 2006-04-17 | 2008-04-30 | 大连理工大学 | Method for expanding hemopoietic stem cell under three-dimensional condition |
US7727763B2 (en) * | 2006-04-17 | 2010-06-01 | Bioe, Llc | Differentiation of multi-lineage progenitor cells to respiratory epithelial cells |
US8435786B2 (en) * | 2006-05-31 | 2013-05-07 | Cellect Biotechnology Ltd. | Methods of selecting stem cells and uses thereof |
JP4851543B2 (en) * | 2007-01-12 | 2012-01-11 | 日本板硝子株式会社 | Three-dimensional cell culture carrier and cell culture method using the same |
US8084023B2 (en) | 2007-01-22 | 2011-12-27 | The Board Of Trustees Of The University Of Arkansas | Maintenance and propagation of mesenchymal stem cells |
US20100172830A1 (en) * | 2007-03-29 | 2010-07-08 | Cellx Inc. | Extraembryonic Tissue cells and method of use thereof |
CA3071055A1 (en) * | 2007-04-07 | 2008-10-16 | Whitehead Institute For Biomedical Research | Reprogramming of somatic cells |
EP2155860B1 (en) * | 2007-05-03 | 2014-08-27 | The Brigham and Women's Hospital, Inc. | Multipotent stem cells and uses thereof |
US8574567B2 (en) * | 2007-05-03 | 2013-11-05 | The Brigham And Women's Hospital, Inc. | Multipotent stem cells and uses thereof |
WO2008152640A2 (en) * | 2007-06-13 | 2008-12-18 | Pluristem Ltd. | Three dimensional biocompatible scaffolds for ex-vivo expansion and transplantation of stem cells |
US20090029463A1 (en) * | 2007-07-25 | 2009-01-29 | Bioe, Inc. | Differentiation of Multi-Lineage Progenitor Cells to Chondrocytes |
PL2200622T5 (en) | 2007-09-19 | 2016-08-31 | Pluristem Ltd | Adherent cells from adipose or placenta tissues and use thereof in therapy |
CN101978045A (en) * | 2007-09-26 | 2011-02-16 | 细胞基因细胞疗法公司 | Angiogenic cells from human placental perfusate |
CA2724839A1 (en) * | 2008-05-21 | 2009-11-26 | Bioe Llc | Differentiation of multi-lineage progenitor cells to pancreatic cells |
US9393273B2 (en) | 2008-05-27 | 2016-07-19 | Pluristem Ltd. | Methods of treating inflammatory colon diseases |
EP3539380A3 (en) | 2008-08-20 | 2019-12-18 | Celularity, Inc. | Improved cell composition and methods of making the same |
CA2734446C (en) * | 2008-08-22 | 2017-06-20 | Anthrogenesis Corporation | Methods and compositions for treatment of bone defects with placental cell populations |
JP2012508577A (en) | 2008-11-12 | 2012-04-12 | カリス ライフ サイエンシズ ルクセンブルク ホールディングス | Method and system for using exosomes to determine phenotype |
RU2562154C2 (en) * | 2008-11-19 | 2015-09-10 | Антродженезис Корпорейшн | Amniotic adhesive cells |
EP3103415B1 (en) | 2009-03-03 | 2020-12-16 | The Trustees of Columbia University in the City of New York | Method for bone tissue engineering using a bioreactor |
WO2010146177A1 (en) * | 2009-06-18 | 2010-12-23 | Mc2 Cell Aps | Bone marrow extracellular matrix extract and therapeutic use thereof |
MX348077B (en) * | 2009-07-20 | 2017-05-25 | Janssen Biotech Inc | Differentiation of human embryonic stem cells. |
NZ600651A (en) | 2009-11-30 | 2014-07-25 | Pluristem Ltd | Adherent cells from placenta and use of same in disease treatment |
US8278101B2 (en) * | 2009-12-07 | 2012-10-02 | Synthecon, Inc. | Stem cell bioprocessing and cell expansion |
CA2791905A1 (en) | 2010-03-01 | 2011-09-09 | Caris Life Sciences Luxembourg Holdings, S.A.R.L. | Biomarkers for theranostics |
EP2556172A4 (en) | 2010-04-06 | 2013-10-30 | Caris Life Sciences Luxembourg Holdings | Circulating biomarkers for disease |
TW201138792A (en) | 2010-04-08 | 2011-11-16 | Anthrogenesis Corp | Treatment of sarcoidosis using placental stem cells |
ES2572211T3 (en) | 2010-05-27 | 2016-05-30 | Pluristem Ltd | Skeletal muscle regeneration using mesenchymal stem cells |
CA2810444A1 (en) | 2010-09-07 | 2012-03-15 | The Board Of Regents Of The University Of Texas System | Tissue-specific differentiation matrices and uses thereof |
US8895291B2 (en) | 2010-10-08 | 2014-11-25 | Terumo Bct, Inc. | Methods and systems of growing and harvesting cells in a hollow fiber bioreactor system with control conditions |
EP2658557A1 (en) | 2010-12-31 | 2013-11-06 | Anthrogenesis Corporation | Enhancement of placental stem cell potency using modulatory rna molecules |
KR101903339B1 (en) | 2011-03-22 | 2018-10-01 | 플루리스템 리미티드 | Methods for treating radiation or chemical injury |
AU2012241521B2 (en) | 2011-04-15 | 2016-07-14 | Pluri Biotech Ltd | Methods and systems for harvesting cells |
WO2012158910A2 (en) * | 2011-05-18 | 2012-11-22 | The Regents Of The University Of California | Compositions and methods for treating retinal diseases |
TWI602570B (en) | 2011-06-01 | 2017-10-21 | 安瑟吉納西斯公司 | Treatment of pain using placental stem cells |
JP6182135B2 (en) | 2011-06-06 | 2017-08-16 | レゲネシス ベーフェーベーアー | Amplification of stem cells in a hollow fiber bioreactor |
JP6095671B2 (en) * | 2011-10-07 | 2017-03-15 | ポール テクノロジー ユーケイ リミテッドPall Technology Uk Limited | Fluid treatment control system and corresponding method |
CA2882687A1 (en) | 2012-09-04 | 2014-03-13 | Pluristem Ltd. | Methods for prevention and treatment of preeclampsia |
US9512393B2 (en) | 2012-09-06 | 2016-12-06 | Pluristem Ltd. | Devices and methods for culture of cells |
US10351910B2 (en) | 2013-02-20 | 2019-07-16 | Pluristem Ltd | Gene and protein expression properties of adherent stromal cells cultured in 3D |
WO2014141111A1 (en) | 2013-03-14 | 2014-09-18 | Pluristem Ltd. | Methods for prevention and treatment of graft-versus-host disease |
KR101442783B1 (en) * | 2013-05-24 | 2014-09-22 | 연세대학교 산학협력단 | Composition for Inducing Erythroid Differentiation |
TWI555847B (en) * | 2013-08-02 | 2016-11-01 | jia-ju Cai | Culture and production of adipose - derived stem cells and their stem cell secretions |
CN105992816B (en) | 2013-11-16 | 2018-04-17 | 泰尔茂比司特公司 | Cell amplification in bioreactor |
CN103690273A (en) * | 2013-11-21 | 2014-04-02 | 中国人民解放军第四军医大学 | In vivo bioreactor |
WO2015132729A1 (en) | 2014-03-04 | 2015-09-11 | Pluristem Ltd. | Systems and methods for growing and harvesting cells |
JP6783143B2 (en) | 2014-03-25 | 2020-11-11 | テルモ ビーシーティー、インコーポレーテッド | Passive replenishment of medium |
EP3129075A4 (en) | 2014-04-10 | 2017-12-06 | Bonus Therapeutics Ltd. | Bone repair compositions |
PL3132247T3 (en) * | 2014-04-16 | 2022-01-03 | Juno Therapeutics Gmbh | Methods, kits and apparatus for expanding a population of cells |
WO2016049421A1 (en) | 2014-09-26 | 2016-03-31 | Terumo Bct, Inc. | Scheduled feed |
WO2016187378A1 (en) * | 2015-05-20 | 2016-11-24 | Celgene Corporation | In vitro cell culture methods for beta-thalassemia using activin type ii receptor ligand traps |
WO2017004592A1 (en) | 2015-07-02 | 2017-01-05 | Terumo Bct, Inc. | Cell growth with mechanical stimuli |
WO2017021543A1 (en) * | 2015-08-05 | 2017-02-09 | Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) | Use of dermatopontin for maintaining hematopoietic stem and/or progenitor cells in culture |
JP7195141B2 (en) | 2015-10-22 | 2022-12-23 | ジュノ セラピューティクス ゲーエムベーハー | Methods, kits, agents and devices for transduction |
MA45488A (en) | 2015-10-22 | 2018-08-29 | Juno Therapeutics Gmbh | CELL CULTURE PROCESSES, KITS AND APPARATUS |
MA45489A (en) | 2015-10-22 | 2018-08-29 | Juno Therapeutics Gmbh | CELL CULTURE PROCESSES, ASSOCIATED KITS AND APPARATUS |
US11104874B2 (en) | 2016-06-07 | 2021-08-31 | Terumo Bct, Inc. | Coating a bioreactor |
US11685883B2 (en) | 2016-06-07 | 2023-06-27 | Terumo Bct, Inc. | Methods and systems for coating a cell growth surface |
DE102016114043B3 (en) * | 2016-07-29 | 2017-08-10 | Technische Universität Dresden | Device for isolating stem cells from fetal tissues |
US20180057784A1 (en) | 2016-08-27 | 2018-03-01 | 3D Biotek, Llc | Bioreactor |
RU2688321C2 (en) * | 2016-12-12 | 2019-05-21 | Федеральное Государственное Бюджетное Учреждение Науки Институт Молекулярной Биологии Им. В.А. Энгельгардта Российской Академии Наук (Имб Ран) | Method for improving the collecting of hematopoietic cells during their cultivation on the stromal layers by pre-magnetization of the latter |
EP3656842A1 (en) | 2017-03-31 | 2020-05-27 | Terumo BCT, Inc. | Cell expansion |
US11624046B2 (en) | 2017-03-31 | 2023-04-11 | Terumo Bct, Inc. | Cell expansion |
IL290068B2 (en) | 2017-04-07 | 2023-09-01 | Epibone Inc | System and method for seeding and culturing |
US11866465B2 (en) | 2017-04-27 | 2024-01-09 | Juno Therapeutics Gmbh | Oligomeric particle reagents and methods of use thereof |
EP3717623A1 (en) * | 2017-11-29 | 2020-10-07 | Corning Incorporated | Filtered cell culture caps and cell culture methods |
US20220033751A1 (en) * | 2018-10-03 | 2022-02-03 | Pluristem Ltd. | Modular Bioreactor |
RU190863U1 (en) * | 2019-02-15 | 2019-07-15 | Федеральное государственное бюджетное учреждение "Российский ордена Трудового Красного Знамени научно-исследовательский институт травматологии и ортопедии имени Р.Р. Вредена" Министерства здравоохранения Российской Федерации (ФГБУ "РНИИТО им. Р.Р. Вредена" Минздрава России) | DEVICE FOR COMPOUND CELLULAR CULTURE AND A Biodegradable Media |
AU2021289205A1 (en) | 2020-06-11 | 2023-02-02 | Mesoblast International Sarl | 3D culture of mesenchymal lineage precursor or stem cells |
WO2022248594A1 (en) | 2021-05-27 | 2022-12-01 | Bühler AG | Optimized industrial bioreactor and method thereof, with mutually dependent, coupled process control loops |
EP4095626A1 (en) | 2021-05-27 | 2022-11-30 | Bühler AG | Optimized industrial bioreactor and method thereof, with mutually dependent, coupled process control loops |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5266476A (en) * | 1985-06-18 | 1993-11-30 | Yeda Research & Development Co., Ltd. | Fibrous matrix for in vitro cell cultivation |
US5437994A (en) * | 1989-06-15 | 1995-08-01 | Regents Of The University Of Michigan | Method for the ex vivo replication of stem cells, for the optimization of hematopoietic progenitor cell cultures, and for increasing the metabolism, GM-CSF secretion and/or IL-6 secretion of human stromal cells |
US5510262A (en) * | 1990-06-18 | 1996-04-23 | Massachusetts Institute Of Technology | Cell-culturing apparatus and method employing a macroporous support |
US5541107A (en) * | 1986-04-18 | 1996-07-30 | Advanced Tissue Sciences, Inc. | Three-dimensional bone marrow cell and tissue culture system |
Family Cites Families (246)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
FR94491E (en) * | 1965-10-13 | 1969-08-22 | Philips Massiot Mat Medic | Pacemaker. |
US6343232B1 (en) | 1966-08-19 | 2002-01-29 | Mower Chf Treatment Irrevocable Trust | Augmentation of muscle contractility by biphasic stimulation |
US6136019A (en) | 1996-08-19 | 2000-10-24 | Mower Family Chf Treatment Irrevocable Trust | Augmentation of electrical conduction and contractility by biphasic cardiac pacing administered via the cardiac blood pool |
NL154600B (en) | 1971-02-10 | 1977-09-15 | Organon Nv | METHOD FOR THE DETERMINATION AND DETERMINATION OF SPECIFIC BINDING PROTEINS AND THEIR CORRESPONDING BINDABLE SUBSTANCES. |
US3587567A (en) | 1968-12-20 | 1971-06-28 | Peter Paul Schiff | Mechanical ventricular assistance assembly |
DE1924227C3 (en) * | 1969-05-12 | 1974-12-05 | Draegerwerk Ag, 2400 Luebeck | Anesthetic vaporizer |
NL154598B (en) | 1970-11-10 | 1977-09-15 | Organon Nv | PROCEDURE FOR DETERMINING AND DETERMINING LOW MOLECULAR COMPOUNDS AND PROTEINS THAT CAN SPECIFICALLY BIND THESE COMPOUNDS AND TEST PACKAGING. |
NL154599B (en) | 1970-12-28 | 1977-09-15 | Organon Nv | PROCEDURE FOR DETERMINING AND DETERMINING SPECIFIC BINDING PROTEINS AND THEIR CORRESPONDING BINDABLE SUBSTANCES, AND TEST PACKAGING. |
US3942536A (en) * | 1971-03-15 | 1976-03-09 | Mieczyslaw Mirowski | Cardioverting device having single intravascular catheter electrode system and method for its use |
US3901654A (en) | 1971-06-21 | 1975-08-26 | Biological Developments | Receptor assays of biologically active compounds employing biologically specific receptors |
US3796221A (en) * | 1971-07-07 | 1974-03-12 | N Hagfors | Apparatus for delivering electrical stimulation energy to body-implanted apparatus with signal-receiving means |
US3853987A (en) | 1971-09-01 | 1974-12-10 | W Dreyer | Immunological reagent and radioimmuno assay |
US3867517A (en) | 1971-12-21 | 1975-02-18 | Abbott Lab | Direct radioimmunoassay for antigens and their antibodies |
NL171930C (en) | 1972-05-11 | 1983-06-01 | Akzo Nv | METHOD FOR DETERMINING AND DETERMINING BITES AND TEST PACKAGING. |
US3796211A (en) * | 1972-08-07 | 1974-03-12 | Medics Res & Dev Inc | Biopsy sampling method and device for the female genital tract |
US3850578A (en) | 1973-03-12 | 1974-11-26 | H Mcconnell | Process for assaying for biologically active molecules |
NL7409823A (en) * | 1973-07-31 | 1975-02-04 | Fujitsu Ltd | OUTPUT DEVICE FOR COORDINATE POSITIONS INFORMATION. |
US3935074A (en) | 1973-12-17 | 1976-01-27 | Syva Company | Antibody steric hindrance immunoassay with two antibodies |
US4316472C1 (en) | 1974-04-25 | 2001-08-14 | Mieczyslaw Mirowski | Cardioverting device with stored energy selecting means and discharge initiating means and related method |
US4572191B1 (en) * | 1974-04-25 | 2000-10-24 | Mirowski Miecyslaw | Command atrial cardioverter |
US3952750A (en) * | 1974-04-25 | 1976-04-27 | Mieczyslaw Mirowski | Command atrial cardioverting device |
US3996345A (en) | 1974-08-12 | 1976-12-07 | Syva Company | Fluorescence quenching with immunological pairs in immunoassays |
US3942750A (en) * | 1974-08-13 | 1976-03-09 | Thomas & Betts Corporation | Adjustable clamp |
US4034074A (en) | 1974-09-19 | 1977-07-05 | The Board Of Trustees Of Leland Stanford Junior University | Universal reagent 2-site immunoradiometric assay using labelled anti (IgG) |
US4202340A (en) * | 1975-09-30 | 1980-05-13 | Mieczyslaw Mirowski | Method and apparatus for monitoring heart activity, detecting abnormalities, and cardioverting a malfunctioning heart |
US4184493A (en) * | 1975-09-30 | 1980-01-22 | Mieczyslaw Mirowski | Circuit for monitoring a heart and for effecting cardioversion of a needy heart |
US4030509A (en) * | 1975-09-30 | 1977-06-21 | Mieczyslaw Mirowski | Implantable electrodes for accomplishing ventricular defibrillation and pacing and method of electrode implantation and utilization |
US3984533A (en) | 1975-11-13 | 1976-10-05 | General Electric Company | Electrophoretic method of detecting antigen-antibody reaction |
US4098876A (en) | 1976-10-26 | 1978-07-04 | Corning Glass Works | Reverse sandwich immunoassay |
US4106494A (en) | 1977-08-29 | 1978-08-15 | American Optical Corporation | Heart defibrillating and monitoring system |
US4164216A (en) | 1978-01-26 | 1979-08-14 | Person Orville W | Throat obstruction expulsion device |
US4223678A (en) | 1978-05-03 | 1980-09-23 | Mieczyslaw Mirowski | Arrhythmia recorder for use with an implantable defibrillator |
US4273114A (en) * | 1978-10-19 | 1981-06-16 | Michigan Instruments, Inc. | Cardiopulmonary resuscitator, defibrillator and monitor |
US4293734A (en) * | 1979-02-23 | 1981-10-06 | Peptek, Incorporated | Touch panel system and method |
US4237895A (en) | 1979-04-20 | 1980-12-09 | Medcor, Inc. | Control signal transmitter and monitor for implanted pacer |
US4403614A (en) | 1979-07-19 | 1983-09-13 | Medtronic, Inc. | Implantable cardioverter |
US4686332A (en) * | 1986-06-26 | 1987-08-11 | International Business Machines Corporation | Combined finger touch and stylus detection system for use on the viewing surface of a visual display device |
US4312354A (en) | 1980-02-04 | 1982-01-26 | Arco Medical Products Company | Pacemaker with circuit for pulse width modulating stimulus pulses in accordance with programmed parameter control states |
US4879219A (en) | 1980-09-19 | 1989-11-07 | General Hospital Corporation | Immunoassay utilizing monoclonal high affinity IgM antibodies |
US4440172A (en) | 1980-10-02 | 1984-04-03 | Mieczyslaw Mirowski | Apparatus for combining pacing and cardioverting functions in a single implanted device |
US4387717A (en) | 1980-10-03 | 1983-06-14 | Research Corporation | Pacer internal cardiac electrogram sensing system |
US4639720A (en) * | 1981-01-12 | 1987-01-27 | Harris Corporation | Electronic sketch pad |
US4407288B1 (en) | 1981-02-18 | 2000-09-19 | Mieczyslaw Mirowski | Implantable heart stimulator and stimulation method |
US4384585A (en) | 1981-03-06 | 1983-05-24 | Medtronic, Inc. | Synchronous intracardiac cardioverter |
US4428366A (en) * | 1981-05-06 | 1984-01-31 | Alfred B. Kurtz | Electromagnetic apparatus and method for the reduction of serum glucose levels |
US4765341A (en) | 1981-06-22 | 1988-08-23 | Mieczyslaw Mirowski | Cardiac electrode with attachment fin |
US4554922A (en) | 1982-09-30 | 1985-11-26 | Prystowsky Eric N | Method of inhibiting cardiac arrhythmias |
DE3244405A1 (en) | 1982-12-01 | 1984-06-07 | Fa. J. Eberspächer, 7300 Esslingen | DEVICE FOR MIXTURE TREATMENT IN MOTOR-INDEPENDENT HEATERS |
CA1199371A (en) * | 1982-12-03 | 1986-01-14 | Orest Z. Roy | Ultrasonic enhancement of cardiac contractility synchronised with ecg event or defibrillation pulse |
US4506680A (en) * | 1983-03-17 | 1985-03-26 | Medtronic, Inc. | Drug dispensing body implantable lead |
US4550221A (en) * | 1983-10-07 | 1985-10-29 | Scott Mabusth | Touch sensitive control device |
US5011771A (en) | 1984-04-12 | 1991-04-30 | The General Hospital Corporation | Multiepitopic immunometric assay |
US4543956A (en) | 1984-05-24 | 1985-10-01 | Cordis Corporation | Biphasic cardiac pacer |
US4628934A (en) | 1984-08-07 | 1986-12-16 | Cordis Corporation | Method and means of electrode selection for pacemaker with multielectrode leads |
US4666828A (en) | 1984-08-15 | 1987-05-19 | The General Hospital Corporation | Test for Huntington's disease |
US4566456A (en) * | 1984-10-18 | 1986-01-28 | Cordis Corporation | Apparatus and method for adjusting heart/pacer rate relative to right ventricular systolic pressure to obtain a required cardiac output |
US4683202A (en) | 1985-03-28 | 1987-07-28 | Cetus Corporation | Process for amplifying nucleic acid sequences |
US4801531A (en) | 1985-04-17 | 1989-01-31 | Biotechnology Research Partners, Ltd. | Apo AI/CIII genomic polymorphisms predictive of atherosclerosis |
US4674508A (en) | 1985-05-28 | 1987-06-23 | Cordis Corporation | Low-power consumption cardiac pacer based on automatic verification of evoked contractions |
US4690155A (en) | 1985-07-03 | 1987-09-01 | Cordis Corporation | Monophasic action potential recording lead |
US4726379A (en) * | 1985-11-14 | 1988-02-23 | Cardiac Control Systems, Inc. | Cardiac pacer with switching circuit for isolation |
US4679572A (en) | 1986-03-11 | 1987-07-14 | Intermedics, Inc. | Low threshold cardiac pacing electrodes |
HU202578B (en) * | 1986-04-18 | 1991-03-28 | Marrow Tech Inc | Process for repricating bone marrow celles in three-dimension celle-cultivating systhem, three-dimension vehicle for the systhem and process for testing citotoxicity of the systhem |
US5032508A (en) * | 1988-09-08 | 1991-07-16 | Marrow-Tech, Inc. | Three-dimensional cell and tissue culture system |
US4830006B1 (en) | 1986-06-17 | 1997-10-28 | Intermedics Inc | Implantable cardiac stimulator for detection and treatment of ventricular arrhythmias |
JPS6370326A (en) * | 1986-09-12 | 1988-03-30 | Wacom Co Ltd | Position detector |
US4726279A (en) * | 1986-11-12 | 1988-02-23 | United Technologies Corporation | Wake stabilized supersonic combustion ram cannon |
DE3732640C1 (en) | 1987-09-28 | 1989-05-18 | Alt Eckhard | Medical device for determining physiological functional parameters |
US5018522A (en) | 1987-10-26 | 1991-05-28 | Medtronic, Inc. | Ramped waveform non-invasive pacemaker |
KR0122737B1 (en) * | 1987-12-25 | 1997-11-20 | 후루다 모또오 | Position detecting device |
US5387419A (en) | 1988-03-31 | 1995-02-07 | The University Of Michigan | System for controlled release of antiarrhythmic agents |
US4914624A (en) * | 1988-05-06 | 1990-04-03 | Dunthorn David I | Virtual button for touch screen |
DE3816042A1 (en) | 1988-05-10 | 1989-11-23 | Alt Eckhard | ENERGY SAVING HEART PACEMAKER |
CA1327838C (en) * | 1988-06-13 | 1994-03-15 | Fred Zacouto | Implantable device to prevent blood clotting disorders |
US5272057A (en) | 1988-10-14 | 1993-12-21 | Georgetown University | Method of detecting a predisposition to cancer by the use of restriction fragment length polymorphism of the gene for human poly (ADP-ribose) polymerase |
JPH0538723Y2 (en) | 1988-12-19 | 1993-09-30 | ||
US4928688A (en) | 1989-01-23 | 1990-05-29 | Mieczyslaw Mirowski | Method and apparatus for treating hemodynamic disfunction |
USRE38119E1 (en) | 1989-01-23 | 2003-05-20 | Mirowski Family Ventures, LLC | Method and apparatus for treating hemodynamic disfunction |
US5031617A (en) | 1989-03-13 | 1991-07-16 | Klettner Harold L | Method of altering human blood glucose levels by the application of electric charge |
US5605822A (en) | 1989-06-15 | 1997-02-25 | The Regents Of The University Of Michigan | Methods, compositions and devices for growing human hematopoietic cells |
US5101814A (en) * | 1989-08-11 | 1992-04-07 | Palti Yoram Prof | System for monitoring and controlling blood glucose |
US5192659A (en) | 1989-08-25 | 1993-03-09 | Genetype Ag | Intron sequence analysis method for detection of adjacent and remote locus alleles as haplotypes |
EP0421025B1 (en) * | 1989-10-02 | 1999-05-06 | Koninklijke Philips Electronics N.V. | Data processing system with a touch screen and a digitizing tablet, both integrated in an input device |
US5402151A (en) * | 1989-10-02 | 1995-03-28 | U.S. Philips Corporation | Data processing system with a touch screen and a digitizing tablet, both integrated in an input device |
US5020544A (en) * | 1989-11-01 | 1991-06-04 | Cardiac Pacemakers, Inc. | Low energy defibrillation electrode |
US5044375A (en) | 1989-12-08 | 1991-09-03 | Cardiac Pacemakers, Inc. | Unitary intravascular defibrillating catheter with separate bipolar sensing |
US4998531A (en) * | 1990-03-28 | 1991-03-12 | Cardiac Pacemakers, Inc. | Implantable N-phasic defibrillator output bridge circuit |
US5097843A (en) | 1990-04-10 | 1992-03-24 | Siemens-Pacesetter, Inc. | Porous electrode for a pacemaker |
US5236413B1 (en) | 1990-05-07 | 1996-06-18 | Andrew J Feiring | Method and apparatus for inducing the permeation of medication into internal tissue |
US5320642A (en) | 1990-06-01 | 1994-06-14 | Board Of Regents For The University Of Ok | Method for alleviating and diagnosing symptoms of heart block |
US5083564A (en) | 1990-06-01 | 1992-01-28 | Board Of Regents Of The University Of Oklahoma | Method for alleviating and diagnosing symptoms of heart block |
EP0486704B1 (en) * | 1990-06-12 | 1999-03-31 | Hitachi Maxell, Ltd. | Organic electrolytic battery |
US5205284A (en) | 1990-06-12 | 1993-04-27 | Zoll Medical Corporation | Method and apparatus for transcutaneous electrical cardiac pacing with background stimulation |
US5499971A (en) * | 1990-06-15 | 1996-03-19 | Cortrak Medical, Inc. | Method for iontophoretically delivering drug adjacent to a heart |
ATE123658T1 (en) | 1990-06-15 | 1995-06-15 | Cortrak Medical Inc | DEVICE FOR DISPENSING MEDICATIONS. |
US5087243A (en) * | 1990-06-18 | 1992-02-11 | Boaz Avitall | Myocardial iontophoresis |
US5156149A (en) | 1990-08-10 | 1992-10-20 | Medtronic, Inc. | Sensor for detecting cardiac depolarizations particularly adapted for use in a cardiac pacemaker |
US5370665A (en) | 1990-08-10 | 1994-12-06 | Medtronic, Inc. | Medical stimulator with multiple operational amplifier output stimulation circuits |
US5584804A (en) | 1990-10-10 | 1996-12-17 | Life Resuscitation Technologies, Inc. | Brain resuscitation and organ preservation device and method for performing the same |
US5163428A (en) | 1990-10-11 | 1992-11-17 | Ventritex, Inc. | Implantable cardiac defibrillator with current leakage detecting means |
US5111815A (en) * | 1990-10-15 | 1992-05-12 | Cardiac Pacemakers, Inc. | Method and apparatus for cardioverter/pacer utilizing neurosensing |
US5137021A (en) | 1990-11-29 | 1992-08-11 | Medtronic, Inc. | Lead current measurement circuit |
US5129394A (en) | 1991-01-07 | 1992-07-14 | Medtronic, Inc. | Method and apparatus for controlling heart rate in proportion to left ventricular pressure |
US5476497A (en) | 1991-01-09 | 1995-12-19 | Ann Mirowski | Oval electrode lead body |
IT1246631B (en) * | 1991-01-25 | 1994-11-24 | Gd Spa | STABILIZATION CONVEYOR OF CIGARETTES PACKAGES OUT OF A PACKAGING MACHINE. |
US5156147A (en) | 1991-02-05 | 1992-10-20 | Cardiac Pacemakers, Inc. | Variable rate pacemaker having upper rate limit governor based on hemodynamic performance |
US5161527A (en) | 1991-02-13 | 1992-11-10 | Telectronics Pacing Systems, Inc. | Apparatus and method for detecting abnormal cardiac rhythms in dual chamber arrhythmia control system |
US5199428A (en) | 1991-03-22 | 1993-04-06 | Medtronic, Inc. | Implantable electrical nerve stimulator/pacemaker with ischemia for decreasing cardiac workload |
US5464020A (en) | 1991-03-25 | 1995-11-07 | Lerner; Albert M. | Diagnosing and treating subacute cardiac dysfunction |
US5482052A (en) | 1991-04-29 | 1996-01-09 | Lerner; Eduard N. | Method and apparatus for determining sensory functions |
WO1994008657A1 (en) | 1992-10-20 | 1994-04-28 | Noel Desmond Gray | A heart pacemaker |
US5168085A (en) | 1991-05-20 | 1992-12-01 | Corning Incorporated | Multi-stage twc system |
US5458568A (en) | 1991-05-24 | 1995-10-17 | Cortrak Medical, Inc. | Porous balloon for selective dilatation and drug delivery |
AU1899292A (en) * | 1991-05-24 | 1993-01-08 | Ep Technologies Inc | Combination monophasic action potential/ablation catheter and high-performance filter system |
US5213098A (en) | 1991-07-26 | 1993-05-25 | Medtronic, Inc. | Post-extrasystolic potentiation stimulation with physiologic sensor feedback |
US5381160A (en) * | 1991-09-27 | 1995-01-10 | Calcomp Inc. | See-through digitizer with clear conductive grid |
ATE142520T1 (en) | 1991-11-04 | 1996-09-15 | Cardiac Pacemakers Inc | IMPLANTABLE HEART MONITORING AND STIMULATION DEVICE FOR DIAGNOSIS AND THERAPY |
US5284491A (en) | 1992-02-27 | 1994-02-08 | Medtronic, Inc. | Cardiac pacemaker with hysteresis behavior |
US5531764A (en) | 1992-03-24 | 1996-07-02 | Angeion Corporation | Implantable defibrillator system and method having successive changeable defibrillation waveforms |
US5342404A (en) | 1992-04-03 | 1994-08-30 | Intermedics, Inc. | Implantable medical interventional device |
US5365461A (en) * | 1992-04-30 | 1994-11-15 | Microtouch Systems, Inc. | Position sensing computer input device |
US5501662A (en) | 1992-05-22 | 1996-03-26 | Genetronics, Inc. | Implantable electroporation method and apparatus for drug and gene delivery |
EP0574213B1 (en) * | 1992-06-08 | 1999-03-24 | Synaptics, Inc. | Object position detector |
US5861583A (en) * | 1992-06-08 | 1999-01-19 | Synaptics, Incorporated | Object position detector |
US6239389B1 (en) * | 1992-06-08 | 2001-05-29 | Synaptics, Inc. | Object position detection system and method |
US5543588A (en) * | 1992-06-08 | 1996-08-06 | Synaptics, Incorporated | Touch pad driven handheld computing device |
US5366486A (en) | 1992-06-25 | 1994-11-22 | Indiana University Foundation | Automatic fibrillation detector and defibrillator apparatus and method |
US5281521A (en) | 1992-07-20 | 1994-01-25 | The Trustees Of The University Of Pennsylvania | Modified avidin-biotin technique |
US5342401A (en) | 1992-08-19 | 1994-08-30 | The Regents Of The University Of California | Real time cardiac arrhythmia stabilizing system |
US5634899A (en) * | 1993-08-20 | 1997-06-03 | Cortrak Medical, Inc. | Simultaneous cardiac pacing and local drug delivery method |
US5320643A (en) | 1992-10-06 | 1994-06-14 | Medtronic, Inc. | Automatic cardiac capture restoration and threshold-seeking method and apparatus |
US5478739A (en) * | 1992-10-23 | 1995-12-26 | Advanced Tissue Sciences, Inc. | Three-dimensional stromal cell and tissue culture system |
US5334222A (en) | 1992-11-03 | 1994-08-02 | Cardiac Pacemakers, Inc. | Cardiac stimulating apparatus and method for heart failure therapy |
SE9203284D0 (en) | 1992-11-04 | 1992-11-04 | Siemens Elema Ab | HJAERTSTIMULATOR |
US5807306A (en) | 1992-11-09 | 1998-09-15 | Cortrak Medical, Inc. | Polymer matrix drug delivery apparatus |
US5346506A (en) | 1992-11-10 | 1994-09-13 | Mower Morton M | Method for establishing defibrillation threshold for a cardiac defibrillator |
US5353800A (en) | 1992-12-11 | 1994-10-11 | Medtronic, Inc. | Implantable pressure sensor lead |
US5327887A (en) | 1993-01-25 | 1994-07-12 | Ludwik Nowakowski | Cardiopulmonary resuscitation device |
US5386837A (en) * | 1993-02-01 | 1995-02-07 | Mmtc, Inc. | Method for enhancing delivery of chemotherapy employing high-frequency force fields |
US5320543A (en) * | 1993-02-04 | 1994-06-14 | Barton Craig S | Flexible plug protector |
US6133906A (en) * | 1993-03-15 | 2000-10-17 | Microtouch Systems, Inc. | Display-integrated stylus detection system |
JP2986047B2 (en) * | 1993-04-29 | 1999-12-06 | インターナショナル・ビジネス・マシーンズ・コーポレイション | Digital input display device and input processing device and method |
US5528002A (en) * | 1993-07-15 | 1996-06-18 | Pentel Kabushiki Kaisha | Noiseproof digitizing apparatus with low power cordless pen |
US5738096A (en) | 1993-07-20 | 1998-04-14 | Biosense, Inc. | Cardiac electromechanics |
US5391199A (en) | 1993-07-20 | 1995-02-21 | Biosense, Inc. | Apparatus and method for treating cardiac arrhythmias |
US5468254A (en) | 1993-07-26 | 1995-11-21 | Cardiac Pacemakers, Inc. | Method and apparatus for defibrillation using a multiphasic truncated exponential waveform |
US5368040A (en) | 1993-08-02 | 1994-11-29 | Medtronic, Inc. | Apparatus and method for determining a plurality of hemodynamic variables from a single, chroniclaly implanted absolute pressure sensor |
US5571997A (en) * | 1993-08-02 | 1996-11-05 | Kurta Corporation | Pressure sensitive pointing device for transmitting signals to a tablet |
BE1007462A3 (en) * | 1993-08-26 | 1995-07-04 | Philips Electronics Nv | Data processing device with touch sensor and power. |
US5443485A (en) | 1993-09-08 | 1995-08-22 | Intermedics, Inc. | Apparatus and method for capture detection in a cardiac stimulator |
AU7729094A (en) * | 1993-09-15 | 1995-04-03 | Pacesetter, Inc. | Synchronized cardioverter shock therapy for preemptive depolarization |
US5415629A (en) * | 1993-09-15 | 1995-05-16 | Henley; Julian L. | Programmable apparatus for the transdermal delivery of drugs and method |
US5476485A (en) | 1993-09-21 | 1995-12-19 | Pacesetter, Inc. | Automatic implantable pulse generator |
US5411531A (en) * | 1993-09-23 | 1995-05-02 | Medtronic, Inc. | Method and apparatus for control of A-V interval |
SE9303736D0 (en) | 1993-11-12 | 1993-11-12 | Siemens Elema Ab | Apparatus intended to sense the physical state of a living being |
US5425363A (en) * | 1993-12-17 | 1995-06-20 | Wang; Yong G. | Plunge electrode for recording multiple intramyocardial monophasic action potential |
US5419763B1 (en) * | 1994-01-04 | 1997-07-15 | Cor Trak Medical Inc | Prostatic drug-delivery catheter |
JPH08205860A (en) * | 1994-01-21 | 1996-08-13 | Usa Government | Method of expanding and transplantation of hematopoietic cell |
US5391192A (en) * | 1994-03-04 | 1995-02-21 | Telectronics Pacing Systems, Inc. | Automatic ventricular pacing pulse threshold determination utilizing an external programmer and a surface electrocardiogram |
IT233201Y1 (en) | 1994-03-24 | 2000-01-26 | Bracco Spa | TWO-COMPONENT DEVICE FOR THE ADMINISTRATION OF DRUGS |
GB9406702D0 (en) * | 1994-04-05 | 1994-05-25 | Binstead Ronald P | Multiple input proximity detector and touchpad system |
KR100300397B1 (en) * | 1994-04-21 | 2001-10-22 | 김순택 | System having touch panel and digitizer function and driving method |
US5562708A (en) | 1994-04-21 | 1996-10-08 | Medtronic, Inc. | Method and apparatus for treatment of atrial fibrillation |
US5540722A (en) | 1994-05-16 | 1996-07-30 | Physiometrix, Inc. | Switch apparatus and method for switching between multiple electrodes for diagnostic and therapeutic procedures |
US5543589A (en) * | 1994-05-23 | 1996-08-06 | International Business Machines Corporation | Touchpad with dual sensor that simplifies scanning |
US5735876A (en) | 1994-05-31 | 1998-04-07 | Galvani Ltd. | Electrical cardiac output forcing method and apparatus for an atrial defibrillator |
JP3186946B2 (en) * | 1994-05-31 | 2001-07-11 | シャープ株式会社 | Coordinate detection device |
US5514162A (en) | 1994-06-07 | 1996-05-07 | Pacesetter, Inc. | System and method for automatically determining the slope of a transfer function for a rate-responsive cardiac pacemaker |
EP0688579B1 (en) | 1994-06-24 | 2001-08-22 | St. Jude Medical AB | Device for heart therapy |
US5601611A (en) | 1994-08-05 | 1997-02-11 | Ventritex, Inc. | Optical blood flow measurement apparatus and method and implantable defibrillator incorporating same |
US5626622A (en) | 1994-09-21 | 1997-05-06 | Telectronics Pacing Systems, Inc. | Dual sensor rate responsive pacemaker |
US5687734A (en) | 1994-10-20 | 1997-11-18 | Hewlett-Packard Company | Flexible patient monitoring system featuring a multiport transmitter |
DE4440386A1 (en) | 1994-11-11 | 1996-05-15 | Pacesetter Ab | Electrodes for medical applications |
TW274598B (en) * | 1994-11-15 | 1996-04-21 | Alps Electric Co Ltd | Coordinate input device for pen of finger tip |
US5622687A (en) * | 1994-11-15 | 1997-04-22 | Molecular Biosystems, Inc. | Calixarene conjugates useful as MRI and CT diagnostic imaging agents |
ATE237676T1 (en) | 1995-02-16 | 2003-05-15 | Forschungszentrum Juelich Gmbh | METHOD FOR CULTIVATION OF ORGAN FUNCTIONAL CELLS |
SE9500620D0 (en) | 1995-02-20 | 1995-02-20 | Pacesetter Ab | Cardiac stimulation device |
JPH08227336A (en) * | 1995-02-20 | 1996-09-03 | Wacom Co Ltd | Pressure sensing mechanism and stylus pen |
US5556421A (en) | 1995-02-22 | 1996-09-17 | Intermedics, Inc. | Implantable medical device with enclosed physiological parameter sensors or telemetry link |
KR100392723B1 (en) * | 1995-02-22 | 2003-11-28 | 코닌클리케 필립스 일렉트로닉스 엔.브이. | Data processing system with input device capable of data input by touch and stylus and input device |
US6041252A (en) | 1995-06-07 | 2000-03-21 | Ichor Medical Systems Inc. | Drug delivery system and method |
US5956020A (en) * | 1995-07-27 | 1999-09-21 | Microtouch Systems, Inc. | Touchscreen controller with pen and/or finger inputs |
US5836311A (en) | 1995-09-20 | 1998-11-17 | Medtronic, Inc. | Method and apparatus for temporarily immobilizing a local area of tissue |
US5782873A (en) | 1995-10-11 | 1998-07-21 | Trustees Of Boston University | Method and apparatus for improving the function of sensory cells |
US5738105A (en) * | 1995-10-24 | 1998-04-14 | Angeion Corporation | Method and apparatus for sensing R-waves using both near field and far field sensing simultaneously |
US6473069B1 (en) * | 1995-11-13 | 2002-10-29 | Cirque Corporation | Apparatus and method for tactile feedback from input device |
US5825352A (en) * | 1996-01-04 | 1998-10-20 | Logitech, Inc. | Multiple fingers contact sensing method for emulating mouse buttons and mouse operations on a touch sensor pad |
CN1168511C (en) | 1996-01-08 | 2004-09-29 | 伊帕斯动力公司 | Electrical muscle controller |
JPH09190268A (en) * | 1996-01-11 | 1997-07-22 | Canon Inc | Information processor and method for processing information |
US5651378A (en) | 1996-02-20 | 1997-07-29 | Cardiothoracic Systems, Inc. | Method of using vagal nerve stimulation in surgery |
US5913876A (en) | 1996-02-20 | 1999-06-22 | Cardiothoracic Systems, Inc. | Method and apparatus for using vagus nerve stimulation in surgery |
US5727569A (en) | 1996-02-20 | 1998-03-17 | Cardiothoracic Systems, Inc. | Surgical devices for imposing a negative pressure to fix the position of cardiac tissue during surgery |
US5683431A (en) | 1996-03-27 | 1997-11-04 | Medtronic, Inc. | Verification of capture by sensing evoked response across cardioversion electrodes |
US5782876A (en) | 1996-04-15 | 1998-07-21 | Medtronic, Inc. | Method and apparatus using windows and an index value for identifying cardic arrhythmias |
US5792198A (en) | 1996-04-30 | 1998-08-11 | Nappholz; Tibor A. | Auto adaptation of RR interval in implantable pacemaker |
US6006134A (en) | 1998-04-30 | 1999-12-21 | Medtronic, Inc. | Method and device for electronically controlling the beating of a heart using venous electrical stimulation of nerve fibers |
US5720768A (en) * | 1996-05-22 | 1998-02-24 | Sulzer Intermedics Inc. | Dual chamber pacing with interchamber delay |
US6128007A (en) * | 1996-07-29 | 2000-10-03 | Motorola, Inc. | Method and apparatus for multi-mode handwritten input and hand directed control of a computing device |
US6411847B1 (en) * | 1996-08-19 | 2002-06-25 | Morton M. Mower | Apparatus for applying cyclic pacing at an average rate just above the intrinsic heart rate |
US6337995B1 (en) | 1996-08-19 | 2002-01-08 | Mower Chf Treatment Irrevocable Trust | Atrial sensing and multiple site stimulation as intervention for atrial fibrillation |
US6178351B1 (en) * | 1996-08-19 | 2001-01-23 | The Mower Family Chf Treatment Irrevocable Trust | Atrial sensing and multiple site stimulation as intervention means for atrial fibrillation |
US6341235B1 (en) * | 1996-08-19 | 2002-01-22 | Mower Chf Treatment Irrevocable Trust | Augmentation of electrical conduction and contractility by biphasic cardiac pacing administered via the cardiac blood pool |
US6141586A (en) | 1996-08-19 | 2000-10-31 | Mower Family Chf Treatment Irrevocable Trust | Method and apparatus to allow cyclic pacing at an average rate just above the intrinsic heart rate so as to maximize inotropic pacing effects at minimal heart rates |
US5871506A (en) | 1996-08-19 | 1999-02-16 | Mower; Morton M. | Augmentation of electrical conduction and contractility by biphasic cardiac pacing |
US6295470B1 (en) | 1996-08-19 | 2001-09-25 | The Mower Family Chf Treatment Irrevocable Trust | Antitachycardial pacing |
US5755740A (en) * | 1996-08-22 | 1998-05-26 | Nappholz; Tibor | Pacemaker with automatic calibration of the response of multiple sensors |
US5713935A (en) * | 1996-08-23 | 1998-02-03 | Sulzer Intermedics Inc. | Method and apparatus for monitored biphasic cardiac impedance sensing |
US5782881A (en) | 1996-09-20 | 1998-07-21 | Lu; Richard | Pacemaker with safety pacing |
US5797967A (en) | 1996-09-27 | 1998-08-25 | Cardiac Pacemakers, Inc. | System and method to reduce defibrillation requirements |
ZA9710342B (en) | 1996-11-25 | 1998-06-10 | Alza Corp | Directional drug delivery stent and method of use. |
US6151586A (en) | 1996-12-23 | 2000-11-21 | Health Hero Network, Inc. | Computerized reward system for encouraging participation in a health management program |
US6086582A (en) | 1997-03-13 | 2000-07-11 | Altman; Peter A. | Cardiac drug delivery system |
US5807234A (en) | 1997-06-27 | 1998-09-15 | Pacesetter, Inc. | Myostimulator control using metabolic demand and muscle performance |
US6440734B1 (en) * | 1998-09-25 | 2002-08-27 | Cytomatrix, Llc | Methods and devices for the long-term culture of hematopoietic progenitor cells |
US6037882A (en) * | 1997-09-30 | 2000-03-14 | Levy; David H. | Method and apparatus for inputting data to an electronic system |
EP1025178B1 (en) * | 1997-10-23 | 2002-12-18 | H.B. Fuller Licensing & Financing, Inc. | Hot melt pressure sensitive adhesive which exhibits minimal staining |
US6392636B1 (en) * | 1998-01-22 | 2002-05-21 | Stmicroelectronics, Inc. | Touchpad providing screen cursor/pointer movement control |
US7663607B2 (en) * | 2004-05-06 | 2010-02-16 | Apple Inc. | Multipoint touchscreen |
US6067470A (en) * | 1998-03-05 | 2000-05-23 | Mower Family Chf Treatment Irrevocable Trust | System and method for multiple site biphasic stimulation to revert ventricular arrhythmias |
US6278443B1 (en) * | 1998-04-30 | 2001-08-21 | International Business Machines Corporation | Touch screen with random finger placement and rolling on screen to control the movement of information on-screen |
JP4142175B2 (en) * | 1998-10-20 | 2008-08-27 | 松下電器産業株式会社 | Graphical user interface device |
MXPA01005267A (en) * | 1998-11-27 | 2002-04-24 | Synaptics Uk Ltd | Position sensor. |
WO2000044018A1 (en) * | 1999-01-26 | 2000-07-27 | Harald Philipp | Capacitive sensor and array |
ES2338405T3 (en) | 1999-02-04 | 2010-05-07 | Pluristem Ltd. | METHOD AND APPARATUS FOR MAINTENANCE AND EXPASION OF HEMOPOYTIC MOTHER CELLS AND / OR PROGENITOR CELLS. |
US6504530B1 (en) * | 1999-09-07 | 2003-01-07 | Elo Touchsystems, Inc. | Touch confirming touchscreen utilizing plural touch sensors |
JP4365493B2 (en) | 1999-10-25 | 2009-11-18 | 東北リコー株式会社 | Printing device |
US6587093B1 (en) * | 1999-11-04 | 2003-07-01 | Synaptics Incorporated | Capacitive mouse |
JP2001142639A (en) * | 1999-11-15 | 2001-05-25 | Pioneer Electronic Corp | Touch panel device |
AU1778001A (en) * | 1999-11-17 | 2001-05-30 | University Of Rochester | Human ex vivo immune system |
US6417846B1 (en) * | 2000-02-02 | 2002-07-09 | Lee Si-Ken | Multifunction input device |
JP2001308247A (en) * | 2000-04-19 | 2001-11-02 | Nec Kansai Ltd | Lead frame and surface mounting type semiconductor device |
US6555235B1 (en) * | 2000-07-06 | 2003-04-29 | 3M Innovative Properties Co. | Touch screen system |
US6690156B1 (en) * | 2000-07-28 | 2004-02-10 | N-Trig Ltd. | Physical object location apparatus and method and a graphic display device using the same |
US6505745B1 (en) * | 2000-08-01 | 2003-01-14 | Richard E Anderson | Holder for articles such as napkins |
US7311905B2 (en) * | 2002-02-13 | 2007-12-25 | Anthrogenesis Corporation | Embryonic-like stem cells derived from post-partum mammalian placenta, and uses and methods of treatment using said cells |
IL156303A0 (en) * | 2000-12-06 | 2004-01-04 | Robert J Hariri | Method of collecting placental stem cells |
US6570557B1 (en) * | 2001-02-10 | 2003-05-27 | Finger Works, Inc. | Multi-touch system and method for emulating modifier keys via fingertip chords |
EP1362095B1 (en) * | 2001-02-14 | 2015-05-27 | Anthrogenesis Corporation | Post-partum mammalian placenta, its use and placental stem cells therefrom |
KR101132545B1 (en) * | 2001-02-14 | 2012-04-02 | 안트로제네시스 코포레이션 | Post-partum mammalian placenta, its use and placental stem cells therefrom |
US6583676B2 (en) * | 2001-06-20 | 2003-06-24 | Apple Computer, Inc. | Proximity/touch detector and calibration circuit |
US6762752B2 (en) * | 2001-11-29 | 2004-07-13 | N-Trig Ltd. | Dual function input device and method |
US20030188899A1 (en) * | 2002-04-09 | 2003-10-09 | Ching-Chuan Chao | Prepositive electro-magnetic tablet with transparent antenna |
US7498171B2 (en) * | 2002-04-12 | 2009-03-03 | Anthrogenesis Corporation | Modulation of stem and progenitor cell differentiation, assays, and uses thereof |
US6875605B1 (en) | 2002-08-21 | 2005-04-05 | Florida State University Research Foundation, Inc. | Modular cell culture bioreactor and associated methods |
KR100459230B1 (en) * | 2002-11-14 | 2004-12-03 | 엘지.필립스 엘시디 주식회사 | touch panel for display device |
CN100538292C (en) * | 2003-02-10 | 2009-09-09 | N-特莱格有限公司 | The touch of Aristogrid detects |
-
2000
- 2000-02-04 ES ES00913340T patent/ES2338405T3/en not_active Expired - Lifetime
- 2000-02-04 RU RU2001124399/13A patent/RU2249039C2/en active
- 2000-02-04 WO PCT/US2000/002688 patent/WO2000046349A1/en not_active Application Discontinuation
- 2000-02-04 CN CNB00806007XA patent/CN100402642C/en not_active Expired - Fee Related
- 2000-02-04 CA CA2360664A patent/CA2360664C/en not_active Expired - Fee Related
- 2000-02-04 DE DE60043534T patent/DE60043534D1/en not_active Expired - Lifetime
- 2000-02-04 AU AU34807/00A patent/AU759719B2/en not_active Ceased
- 2000-02-04 BR BR0009403-0A patent/BR0009403A/en not_active Application Discontinuation
- 2000-02-04 EP EP00913340A patent/EP1147176B1/en not_active Expired - Lifetime
- 2000-02-04 US US09/890,401 patent/US6911201B1/en not_active Expired - Lifetime
- 2000-02-04 EP EP10184233.4A patent/EP2311938B1/en not_active Expired - Lifetime
- 2000-02-04 AT AT00913340T patent/ATE452181T1/en not_active IP Right Cessation
- 2000-02-04 EP EP09174118.1A patent/EP2208782B1/en not_active Expired - Lifetime
- 2000-02-04 MX MXPA01007820A patent/MXPA01007820A/en not_active IP Right Cessation
- 2000-02-04 JP JP2000597409A patent/JP4523169B2/en not_active Expired - Fee Related
- 2000-02-04 KR KR1020017009869A patent/KR20020013496A/en not_active Application Discontinuation
- 2000-02-04 NZ NZ513303A patent/NZ513303A/en not_active IP Right Cessation
-
2001
- 2001-07-30 IL IL144629A patent/IL144629A/en not_active IP Right Cessation
- 2001-08-07 ZA ZA200106483A patent/ZA200106483B/en unknown
-
2002
- 2002-10-24 HK HK02107728.2A patent/HK1046154B/en not_active IP Right Cessation
-
2005
- 2005-04-11 US US11/102,623 patent/US7678573B2/en not_active Expired - Fee Related
- 2005-04-11 US US11/102,654 patent/US20050180958A1/en not_active Abandoned
- 2005-04-11 US US11/102,635 patent/US20050176137A1/en not_active Abandoned
- 2005-04-11 US US11/102,625 patent/US7534609B2/en not_active Expired - Lifetime
-
2008
- 2008-09-02 US US12/230,566 patent/US20090004738A1/en not_active Abandoned
-
2012
- 2012-01-27 US US13/360,068 patent/US20120122220A1/en not_active Abandoned
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5266476A (en) * | 1985-06-18 | 1993-11-30 | Yeda Research & Development Co., Ltd. | Fibrous matrix for in vitro cell cultivation |
US5541107A (en) * | 1986-04-18 | 1996-07-30 | Advanced Tissue Sciences, Inc. | Three-dimensional bone marrow cell and tissue culture system |
US5437994A (en) * | 1989-06-15 | 1995-08-01 | Regents Of The University Of Michigan | Method for the ex vivo replication of stem cells, for the optimization of hematopoietic progenitor cell cultures, and for increasing the metabolism, GM-CSF secretion and/or IL-6 secretion of human stromal cells |
US5510262A (en) * | 1990-06-18 | 1996-04-23 | Massachusetts Institute Of Technology | Cell-culturing apparatus and method employing a macroporous support |
Cited By (46)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2208782A3 (en) * | 1999-02-04 | 2010-11-03 | Pluristem Ltd. | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells |
WO2000069449A3 (en) * | 1999-05-14 | 2001-02-08 | Advanced Tissue Sciences Inc | Conditioned cell culture medium compositions and methods of use |
US9458486B2 (en) | 1999-05-14 | 2016-10-04 | Allergen, Inc. | Conditioned cell culture medium compositions and methods of use |
US6372494B1 (en) | 1999-05-14 | 2002-04-16 | Advanced Tissue Sciences, Inc. | Methods of making conditioned cell culture medium compositions |
WO2000069449A2 (en) * | 1999-05-14 | 2000-11-23 | Advanced Tissue Sciences, Inc. | Conditioned cell culture medium compositions and methods of use |
US8476231B2 (en) | 1999-05-14 | 2013-07-02 | Allergan, Inc. | Conditioned cell culture medium compositions and methods of use |
US8361485B2 (en) | 1999-05-14 | 2013-01-29 | Skinmedica, Inc. | Conditioned cell culture medium compositions and methods of use |
US8138147B2 (en) | 1999-05-14 | 2012-03-20 | Skinmedica, Inc. | Conditioned cell culture medium compositions and methods of use |
GB2366295A (en) * | 2000-05-10 | 2002-03-06 | Tristem Ireland Ltd | Device and methods to prepare undifferentiated cells |
GB2366295B (en) * | 2000-05-10 | 2003-09-10 | Tristem Ireland Ltd | A device and methods for preparing undifferentiated cells |
WO2002024874A3 (en) * | 2000-09-21 | 2003-02-13 | Schering Corp | Methods for preparing interferon producing dentitric cells |
US6913924B2 (en) | 2000-09-21 | 2005-07-05 | Schering Corporation | Population of interferon producing cells |
US6660523B2 (en) | 2000-09-21 | 2003-12-09 | Schering Corporation | Dendritic cells; methods |
WO2002024874A2 (en) * | 2000-09-21 | 2002-03-28 | Schering Corporation | Methods for preparing interferon producing dentitric cells |
US7160726B2 (en) | 2001-06-07 | 2007-01-09 | Skin Medica, Inc. | Compositions comprising conditioned cell culture media and uses thereof |
US10723986B2 (en) | 2002-04-08 | 2020-07-28 | Octane Biotech Inc. | Automated tissue engineering system |
US9701932B2 (en) | 2002-04-08 | 2017-07-11 | Octane Biotech Inc. | Automated tissue engineering system |
US10844338B1 (en) | 2002-04-08 | 2020-11-24 | Octane Biotech Inc. | Automated tissue engineering system |
US9534195B2 (en) | 2002-04-08 | 2017-01-03 | Octane Biotech Inc. | Automated tissue engineering system |
US8492140B2 (en) | 2002-04-08 | 2013-07-23 | Octane Biotech Inc. | Automated tissue engineering system |
WO2003104386A1 (en) * | 2002-05-22 | 2003-12-18 | 株式会社エムビーエス | Culture apparatus, artificial tissue and blood preparation |
US8846393B2 (en) | 2005-11-29 | 2014-09-30 | Gamida-Cell Ltd. | Methods of improving stem cell homing and engraftment |
EP3091071B1 (en) | 2006-03-23 | 2019-07-03 | Pluristem Ltd. | Methods for cell expansion and uses of cells and conditioned media produced thereby for therapy |
EP2626417A1 (en) * | 2006-03-23 | 2013-08-14 | Pluristem Ltd. | Methods for cell expansion and uses of cells and conditioned media produced thereby for therapy |
EP3091071A1 (en) * | 2006-03-23 | 2016-11-09 | Pluristem Ltd. | Methods for cell expansion and uses of cells and conditioned media produced thereby for therapy |
EP2548951A1 (en) * | 2006-03-23 | 2013-01-23 | Pluristem Ltd. | Methods for cell expansion and uses of cells and conditioned media produced thereby for therapy |
EP2366775A1 (en) * | 2006-03-23 | 2011-09-21 | Pluristem Ltd. | Methods for cell expansion and uses of cells and conditioned media produced thereby for therapy |
EP2010647A4 (en) * | 2006-03-23 | 2010-05-19 | Pluristem Ltd | Methods for cell expansion and uses of cells and conditioned media produced thereby for therapy |
EP2010647A2 (en) * | 2006-03-23 | 2009-01-07 | Pluristem Ltd. | Methods for cell expansion and uses of cells and conditioned media produced thereby for therapy |
US9758762B2 (en) | 2008-09-02 | 2017-09-12 | Pluristem Ltd. | Perfusion bioreactor for culturing CD200—placenta adherent cells |
US10047345B2 (en) | 2012-02-13 | 2018-08-14 | Gamida-Cell Ltd. | Culturing of mesenchymal stem cells with FGF4 and nicotinamide |
US9175266B2 (en) | 2012-07-23 | 2015-11-03 | Gamida Cell Ltd. | Enhancement of natural killer (NK) cell proliferation and activity |
US9567569B2 (en) | 2012-07-23 | 2017-02-14 | Gamida Cell Ltd. | Methods of culturing and expanding mesenchymal stem cells |
US10508264B2 (en) | 2015-01-26 | 2019-12-17 | Ube Industries, Ltd. | Cell culture method using bone marrow-like structure, and porous polyimide film for healing bone injury site |
US10590388B2 (en) | 2015-01-26 | 2020-03-17 | Ube Industries, Ltd. | Cell culture method using bone marrow-like structure, and porous polyimide film for healing bone injury site |
EP3406272A1 (en) | 2015-01-26 | 2018-11-28 | Ube Industries, Ltd. | Cell culture method using bone marrow-like structure, and porous polymide film for healing bone injury site |
EP3464565A4 (en) * | 2016-05-25 | 2020-01-01 | Terumo BCT, Inc. | Cell expansion |
US11371018B2 (en) | 2017-09-01 | 2022-06-28 | Octane Biotech Inc. | End-to-end cell therapy automation |
US11447745B2 (en) | 2017-09-01 | 2022-09-20 | Lonza Walkersville, Inc. | End-to-end cell therapy automation |
US11781113B2 (en) | 2017-09-01 | 2023-10-10 | Lonza Walkersville, Inc. | End-to-end cell therapy automation |
US11827902B2 (en) | 2017-09-01 | 2023-11-28 | Lonza Walkersville, Inc. | End-to-end cell therapy automation |
US11714096B2 (en) | 2018-12-21 | 2023-08-01 | Octane Biotech Inc. | Carousel for modular biologic production units |
US11718833B2 (en) | 2018-12-21 | 2023-08-08 | Lonza Walkersville, Inc. | Automated production of viral vectors |
US11597905B2 (en) | 2018-12-28 | 2023-03-07 | Octane Biotech Inc. | Cell culture and tissue engineering systems with controlled environmental zones |
US11773365B2 (en) | 2019-02-08 | 2023-10-03 | Lonza Walkersville, Inc. | Cell concentration methods and devices for use in automated bioreactors |
WO2021110908A1 (en) | 2019-12-04 | 2021-06-10 | Centre Hospitalier Universitaire Vaudois (C.H.U.V.) | Device and process for tissue-engineering and regenerative medicine |
Also Published As
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CA2360664C (en) | Method and apparatus for maintenance and expansion of hemopoietic stem cells and/or progenitor cells | |
US20100233130A1 (en) | Method and Apparatus for Maintenance and Expansion of Hematopoietic Stem Cells From Mononuclear Cells | |
Bagley et al. | Extended culture of multipotent hematopoietic progenitors without cytokine augmentation in a novel three-dimensional device | |
US8278101B2 (en) | Stem cell bioprocessing and cell expansion | |
EP1627914A2 (en) | Methods and devices for the long-term culture of hematopoietic progenitor cells | |
AU778504B2 (en) | Human brain endothelial cells and growth medium and method for expansion of primitive CD34+CD38- bone marrow stem cells | |
WO2008152640A2 (en) | Three dimensional biocompatible scaffolds for ex-vivo expansion and transplantation of stem cells | |
LaIuppa | Defined culture conditions for ex vivo expansion of megakaryocytes and myeloid progenitors |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
WWE | Wipo information: entry into national phase |
Ref document number: 00806007.X Country of ref document: CN |
|
AK | Designated states |
Kind code of ref document: A1 Designated state(s): AE AL AM AT AU AZ BA BB BG BR BY CA CH CN CR CU CZ DE DK DM EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX NO NZ PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZW |
|
AL | Designated countries for regional patents |
Kind code of ref document: A1 Designated state(s): GH GM KE LS MW SD SL SZ TZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG |
|
121 | Ep: the epo has been informed by wipo that ep was designated in this application | ||
DFPE | Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101) | ||
WWE | Wipo information: entry into national phase |
Ref document number: 144629 Country of ref document: IL |
|
WWE | Wipo information: entry into national phase |
Ref document number: 09890401 Country of ref document: US |
|
WWE | Wipo information: entry into national phase |
Ref document number: 513303 Country of ref document: NZ |
|
WWE | Wipo information: entry into national phase |
Ref document number: PA/a/2001/007820 Country of ref document: MX |
|
ENP | Entry into the national phase |
Ref document number: 2360664 Country of ref document: CA Ref document number: 2360664 Country of ref document: CA Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 34807/00 Country of ref document: AU |
|
WWE | Wipo information: entry into national phase |
Ref document number: 1020017009869 Country of ref document: KR |
|
ENP | Entry into the national phase |
Ref document number: 2000 597409 Country of ref document: JP Kind code of ref document: A |
|
WWE | Wipo information: entry into national phase |
Ref document number: 200106483 Country of ref document: ZA |
|
WWE | Wipo information: entry into national phase |
Ref document number: 2000913340 Country of ref document: EP Ref document number: IN/PCT/2001/1131/CHE Country of ref document: IN |
|
WWP | Wipo information: published in national office |
Ref document number: 2000913340 Country of ref document: EP |
|
REG | Reference to national code |
Ref country code: DE Ref legal event code: 8642 |
|
WWP | Wipo information: published in national office |
Ref document number: 1020017009869 Country of ref document: KR |
|
WWG | Wipo information: grant in national office |
Ref document number: 34807/00 Country of ref document: AU |
|
WWW | Wipo information: withdrawn in national office |
Ref document number: 1020017009869 Country of ref document: KR |