CA2239815A1 - A method and apparatus for holding cells - Google Patents

A method and apparatus for holding cells

Info

Publication number
CA2239815A1
CA2239815A1 CA002239815A CA2239815A CA2239815A1 CA 2239815 A1 CA2239815 A1 CA 2239815A1 CA 002239815 A CA002239815 A CA 002239815A CA 2239815 A CA2239815 A CA 2239815A CA 2239815 A1 CA2239815 A1 CA 2239815A1
Authority
CA
Canada
Prior art keywords
cells
wells
biochamber
cell
stem cells
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CA002239815A
Other languages
French (fr)
Other versions
CA2239815C (en
Inventor
Raymond K. Houck
Joel S. Greenberger
Paul A. Dimilla
Michael M. Domach
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Pittsburgh
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Family has litigation
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Application filed by Individual filed Critical Individual
Publication of CA2239815A1 publication Critical patent/CA2239815A1/en
Application granted granted Critical
Publication of CA2239815C publication Critical patent/CA2239815C/en
Anticipated expiration legal-status Critical
Expired - Lifetime legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • 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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/12—Well or multiwell plates
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
    • C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/48—Automatic or computerized control
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/16—Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/802—Logarithmic growth phase
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/808—Optical sensing apparatus
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/809—Incubators or racks or holders for culture plates or containers
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/813—Continuous fermentation

Abstract

The present invention pertains to an apparatus for holding cells (200). The apparatus comprises a mechanism for incubating cells having a dynamically controlled environment in which the cells are grown, which are maintained in a desired condition and in which cells can be examined while the environment is dynamically controlled and maintained in the desired condition. The apparatus also comprises a mechanism for determining the state of the cells. The determining mechanism is in communication with the incubating mechanism. The present invention pertains to a method of holding cells. The method comprises the steps of incubating the cells in a dynamically controlled environment which is maintained in a desired condition and in which the cells can be examined while the environment is dynamically controlled and maintained in the desired condition. Additionally, there is the step of determining the state of the cells.

Claims (46)

1. An apparatus for holding cells comprising:

a mechanism for incubating cells having a dynamically controlled environment in which the cells are grown, which is maintained in a desired condition and in which cells can be examined while the environment is dynamically controlled and maintained in the desired condition; and a mechanism for determining the state of the cells, said determining mechanism in communication with the incubating mechanism.
2. An apparatus as described in Claim 1 wherein the incubating mechanism includes a housing having a biochamber in the housing.
3. An apparatus as described in Claim 2 wherein the incubating mechanism includes a first well and at least a second well in which cells are grown, said first and second well disposed in the biochamber of the housing.
4. An apparatus as described in Claim 3 wherein the housing has a first port mechanism through which the first and second wells in the biochamber can be viewed, and the determining mechanism includes an imaging mechanism disposed adjacent the first port mechanism which images the cells in the first and second wells.
5. An apparatus as described in Claim 4 wherein the housing has a second port mechanism in fluid communication with the biochamber, and the incubating mechanism includes a mechanism for controlling the environment in the biochamber, said environment controlling mechanism connected with the second port mechanism.
6. An apparatus as described in Claim 5 wherein the environment controlling mechanism includes a heating mechanism in thermal communication with the biochamber to maintain the cells in the first and second wells at a desired temperature.
7. An apparatus as described in Claim 6 wherein the imaging mechanism comprises a computer for identifying whether a cell in the first well or the second well has multiplied, said computer connected to the imaging mechanism to receive images of the first and second wells from the imaging mechanism.
8. An apparatus as described in Claim 7 wherein the imaging mechanism comprises a microscope mechanism which views the first and the second wells, said microscope mechanism disposed adjacent the first port mechanism, said microscope mechanism in communication with the computer.
9. An apparatus as described in Claim 8 wherein the imaging mechanism comprises a camera mechanism for imaging the cells in the first and second wells, said camera mechanism connected to the microscope mechanism such that the camera mechanism takes images of the cells in the first and second wells through the microscope mechanism, said camera mechanism connected to the computer.
10. An apparatus as described in Claim 9 wherein the determining mechanism includes a moving mechanism for moving the first and second wells relative to the microscope mechanism so the microscope mechanism can view the cells in the first and second wells.
11. An apparatus as described in Claim 10 wherein the first port mechanism includes a first window disposed in the top of the housing and a second window disposed in the bottom of the housing and in optical alignment with the first window to form an optical path for light to pass between the first window and the second window.
12. An apparatus as described in Claim 11 wherein the incubating mechanism comprises a transparent plate in which the first and second wells are disposed.
13. An apparatus as described in Claim 12 wherein the environment controlling mechanism comprises a mechanism for controlling media pH in the first and second wells in communication with the biochamber, and a mechanism for controlling pressure in the biochamber in communication with the biochamber.
14. An apparatus as described in Claim 13 wherein the incubating mechanism includes a robotic mechanism for automatically dispensing and aspirating media to and from the first or second wells, said robotic mechanism disposed adjacent the biochamber in movable contact with the first and second wells.
15. An apparatus as described in Claim 14 wherein the robotic mechanism includes a reservoir mechanism for fresh and waste media regarding the first and second wells.
16. An apparatus as described in Claim 15 wherein the determining mechanism includes a diagnostic mechanism in communication with the robotic mechanism for ascertaining an occurrence of a predetermined biological event in the first or second wells.
17. An apparatus as described in Claim 16 wherein the determining mechanism includes a joystick connected to the microscope mechanism to control the position of the microscope mechanism relative to the first and second wells.
18. An apparatus as described in Claim 17 wherein the moving mechanism includes a translation system which moves the plate to align the first or second well with the microscope mechanism.
19. An apparatus as described in Claim 18 wherein the robotic mechanism includes a z-robot pipette through which media is aspirated from or dispensed into the first or second wells.
20. An apparatus as described in Claim 19 wherein the imaging mechanism includes fluorescent material which is introduced to the first or second wells by the z-robot pipette to enhance the imaging of cells in the first or second wells.
21. An apparatus as described in Claim 20 wherein the robotic mechanism comprises a syringe pump mechanism connected to the z-robot pipette and the reservoir mechanism for delivering liquid to the z-robot pipette from the reservoir mechanism.
22. An apparatus as described in Claim 21 wherein the syringe pump mechanism comprises a stepper motor mechanism which provides force to deliver the liquid.
23. An apparatus as described in Claim 22 wherein the reservoir mechanism includes a first reservoir with the growth medium and at least a second reservoir with the quiescence medium.
24. An apparatus as described in Claim 23 wherein the reservoir mechanism includes a distribution valve mechanism connected to the syringe pump mechanism, the z-robot pipette and the first and second reservoirs for directing which liquid from which reservoir is delivered to the z-robot pipette.
25. An apparatus as described in Claim 24 wherein the z-robot pipette has a sensor mechanism which senses the presence of fluid about the tip of the pipette and produces a pipette signal corresponding to the presence of fluids about the tip.
26. An apparatus as described in Claim 25 including a computer which receives the pipette signal, said computer connected to the stepper motor to control the operation of the stepper motor.
27. An apparatus is described in Claim 26 wherein the housing has a bulkhead with a hole through which the z-robot pipette extends into the biochamber.
28. An apparatus as described in Claim 27 wherein the distribution valve mechanism is connected to the diagnostic mechanism to receive medium from the first or second wells.
29. An apparatus as described in Claim 28 wherein the distribution valve mechanism is connected to the computer which controls the distribution valve mechanism.
30. An apparatus as described in Claim 29 wherein the diagnostic mechanism includes an assay for analyzing the media.
31. An apparatus as described in Claim 30 wherein the assay includes a column.
32. An apparatus as described in Claim 31 wherein the assay includes a chromatograph.
33. An apparatus for managing cells comprising:

a housing having a biochamber and at least a first well in the biochamber in which the cells are disposed; and a robotic mechanism for automatically dispensing and aspirating media to and from the first well, said robotic mechanism in contact with the biochamber.
34. A method for holding cells in a chamber comprising the steps of:

scanning a first cell; and determining when the first cell has divided.
35. A method as described in Claim 34 including before the scanning step, there is the step of placing growth medium with the first cell.
36. A method as described in Claim 35 including before the scanning step, there is the step of identifying the location of the first cell.
37. A method as described in Claim 36 including after the determining step, there is the step of replacing growth medium with quiescence medium.
38. A method as described in Claim 37 including before the scanning step, there is the step of placing growth medium with a second cell, and identifying the location of the second cell.
39. A method as described in Claim 38 including after the scanning step there is the step of scanning the second cell.
40. A method as described in Claim 39 wherein the replacing step includes the steps of moving a z-robot pipette into a position to aspirate growth medium from about the first cell; aspirating the growth medium from the about the first cell, and adding quiescence medium about the first cell.
41. A method for using stem cells comprising the steps of:

obtaining stem cells;

placing the stem cells in an automated system for growing stem cells;

growing additional stem cells in the automated system from the stem cells placed in the automated system;
and gathering the stem cells in the automated system.
42. A method as described in Claim 41 including after the gathering step, there is the step of transplanting stem cells into a patient.
43. A method as described in Claim 42 wherein the obtaining step includes the step of removing the stem cells from the patient.
44. A method as described in Claim 43 wherein the growing step includes the step of maximizing growing pluripotent stem cells while minimizing growing differentiated mature blood cells.
45. A method of producing stem cells comprising the steps of:

placing growth medium with a first stem sell in situ;

determining when the first stem cell divides into a second and a third stem cell in situ; and exchanging the growth medium about the second and third stem cells in situ with quiescent medium in situ which inhibits differentiation of the second and third stem cells into committed progenitors.
46. A method for holding cells comprising the steps of:

incubating the cells in a dynamically controlled environment which is maintained in a desired condition and in which the cells can be examined while the environment is dynamically controlled and maintained in the desired condition; and determining the state of the cells.
CA2239815A 1996-11-01 1997-10-31 A method and apparatus for holding cells Expired - Lifetime CA2239815C (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US08/741,628 1996-11-01
US08/741,628 US6008010A (en) 1996-11-01 1996-11-01 Method and apparatus for holding cells
PCT/US1997/019834 WO1998020108A1 (en) 1996-11-01 1997-10-31 A method and apparatus for holding cells

Publications (2)

Publication Number Publication Date
CA2239815A1 true CA2239815A1 (en) 1998-05-14
CA2239815C CA2239815C (en) 2011-10-25

Family

ID=24981507

Family Applications (1)

Application Number Title Priority Date Filing Date
CA2239815A Expired - Lifetime CA2239815C (en) 1996-11-01 1997-10-31 A method and apparatus for holding cells

Country Status (7)

Country Link
US (6) US6008010A (en)
EP (1) EP0877792B1 (en)
JP (2) JP4099236B2 (en)
AU (1) AU742909B2 (en)
CA (1) CA2239815C (en)
DE (1) DE69740113D1 (en)
WO (1) WO1998020108A1 (en)

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