WO2006122088A1

WO2006122088A1 - Cell culture apparatus

Info

Publication number: WO2006122088A1
Application number: PCT/US2006/017848
Authority: WO
Inventors: Vaughan Clift; Tiffany Conerly; Scott Senner; Laura Licato
Original assignee: Isolagen Technologies, Inc.
Priority date: 2005-05-09
Filing date: 2006-05-08
Publication date: 2006-11-16

Abstract

A container (102) includes an upper portion (104) and a lower portion (106). A first surface is inside the container and defines a first passage (108) between the upper portion and the lower portion. A second surface inside the container defines a second passage (112) between the lower portion and the upper portion. A mechanism (122) is adapted to motivate fluid flow from the lower portion to the upper portion through the second passage. A portion of the first surface is adapted for growing cells thereupon.

Description

CELL CULTURE APPARATUS

BACKGROUND

Cells can be taken from a living organism and grown (cultured) under controlled, sterile conditions.

In some instances, cell samples for culturing are obtained from human patients. Those cell samples can be placed into a sterile cell culture apparatus, in which the cells increase in number exponentially. Indeed, it can be possible to generate hundreds of millions of new living cells from one original sample of a few million or even less, over a period of time. Those new living cells can be harvested from the cell culture apparatus and, for example, injected into the patient from which the original sample was taken.

Cultured cells are useful in treating a variety of conditions.

SUMMARY

One aspect features a cell culture apparatus includes a container with an upper portion and a lower portion therein. First surfaces inside the container define a first passage between the upper portion and the lower portion. At least a portion of those first surfaces is adapted for growing cells thereupon. Second surfaces inside the container define a second passage between the lower portion and the upper portion. A mechanism is adapted to motivate fluid flow from the lower portion to the upper portion through the second passage.

In some implementations, the first passage includes multiple plates aligned in a stacked fashion between the upper portion and the lower portion and at least one spacer positioned between adjacent plates. The spacers define open spaces between adjacent plates. In those instances, each plate includes at least one aperture. The first passage includes at least one of the apertures through each plate and at least a portion of the space between adjacent plates. The plates are arranged in such a manner that the apertures of adjacent plates are not aligned with each other. Certain embodiments include a container that is cylindrical, each plate and each spacer is disk-shaped. Each aperture is rectangular in shape. Each aperture originates at a peripheral surface of the disk-shaped plate and extends inward to a position proximate a center point of an associated plate. Indeed, each plate can include a plurality of such apertures extending outward along respective straight lines each of which is positioned to extend from the center point in a direction that is angularly displaced from a straight line associated with an adjacent aperture on the same plate. The space between adjacent plates permits fluid communication between a first aperture of an upper plate and a second aperture of a lower adjacent plate. The surface for growing cells thereupon comprises at least a portion of the space between adjacent plates. The first aperture and the second aperture of a single first passage are not vertically aligned with each other.

According to some implementations, the cell culture apparatus includes a support element positioned inside the container and having support shelves to support the plurality of plates and the at least one spacer. The second passage is positioned inside the support element.

In certain embodiments, the mechanism adapted to motivate fluid flow is an air lift pump operationally coupled to an entry port of the second passage. The entry port is located proximate a lower area of the container. A mechanism adapted to motivate fluid flow can include a heating element adapted to heat fluid located proximate an entry port of the second passage located proximate the lower area of the container.

The first passage is generally arranged so as to accommodate laminar flow of fluid there through. In some implementations, the second passage is positioned at least partially outside the container.

Certain embodiments include an impedance sensor coupled to the surface for growing cells. The impedance sensor is adapted to measure impedance associated with the surface for growing cells. In some instances, a heating element is operationally coupled to the container to heat fluid inside the container. A control unit is adapted to modulate a rate of cell growth in the container by modulating a temperature of the fluid inside the container with the heating element. In some instances, a light source is operationally coupled to the container and adapted to transmit light into the fluid inside the container. A controller can be adapted to control operation of the light source to modulate a rate of cell growth inside the container.

According to another aspect, a method of growing cells includes providing an initiated cell sample on a surface that is preferably tissue culture treated and flowing media over the cell sample in a substantially laminar manner. In one implementation, the method includes removing or harvesting the original cell sample and newly grown cells from the tissue culture treated surface by flowing a proteolytic enzyme (e.g. Trypsin) and/or a metal-chelator (e.g., ethylenediaminetetraacetic (EDTA)) over the cells in a substantially laminar manner.

In some implementations, one or more of the following advantages can be present. The amount of cell growth that can be realized on a given surface area can be improved. Fluid flowing in a laminar manner over the cell growth surfaces can more efficiently deliver nutrients and oxygen to the cells that are growing on those surfaces. The laminar flow can also expose the growing cells to substantially continuous sheer force. This sheer force can facilitate improved growth. The ease and efficiency of collecting cultured cells from a cell culture device can be improved. The area of cell growing surface that can be accommodated in a given volume of space can be increased.

A closed cell growth system, including a source of initiated cells, a culture apparatus, a harvesting apparatus, etc., can be created. An advantage of such a closed system is that the possibility of microbial (e.g., bacterial or fungal) contamination of the culture and/or cells obtained from the culture can be reduced.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a cutaway view of a cell culture apparatus.

FIG. 2 is an exploded view of plates and spacers that might be positioned inside a cell culture apparatus container. FIG. 3 is a cutaway view of an alternative cell culture apparatus.

FIG. 4 is an isometric view of spiral surfaces that might be positioned inside a cell culture apparatus container.

FIG. 5 is a cutaway view of yet another alternative cell culture apparatus. FIG. 6 is a system block diagram.

FIG. 7 is a system piping diagram.

FIG. 8 is a connection diagram for a cell culture apparatus. Like reference numerals refer to similar elements.

DETAILED DESCRIPTION

FIG. 1 discloses a particular implementation of a cell culture apparatus 100 that includes a closed container 102 with an upper portion 104 and a lower portion 106 therein. Surfaces inside the container 102 define a first passage, indicated by a dashed line 108, between the upper portion 104 and the lower portion 106 of the container 102. At least some of those surfaces are adapted to facilitate the growth of cells thereupon. Other surfaces inside the container define a second passage 112 between the lower portion 106 and the upper portion 104 of the container 102. A pumping mechanism 114 is provided to motivate fluid flow from the lower portion 106 to the upper portion 104 via the second passage 112. According to one embodiment, cultures are initiated by adding cells suspended in a culture medium to a cell culture apparatus 100 via a connection on the cell culture apparatus 100. The cells settle onto, and adhere to the those surfaces 11 Oa-11Oe of the first passage 108 adapted to facilitate cell growth thereupon. The cells can be any mammalian cells and will preferably be cells that adhere to the cell growing surfaces in the apparatus. Such cells include, without limitation, fibroblasts, epithelial cells (e.g., keratinocytes), endothelial cells, neuronal cells, adipose tissue cells, muscle cells, leukocytes (e.g., macrophages, monocytes and granulocytes). Subjects from which the cells are obtained include, without limitation, any mammal, including humans and non-human primates (e.g., a monkey, a chimpanzee, or a baboon), horses, bovine animals, pigs, sheep, goats, rabbits, guinea pigs, gerbils, hamsters, rats or mice.

As illustrated, the container 102 is partially filled with fluid, such as tissue culture medium. A fluid level in the container 102 is indicated by reference numeral 130. The pumping mechanism 114 motivates fluid to flow from the lower portion 106 of the container 102 to the upper portion 104 of the container 102 via the second passage 112. Fluid also flows in a substantially laminar manner from the upper portion 104 of the container 102 to the lower portion 106 of the container via the first passage 108. As the fluid flows downward through the first passage 108 it passes the cells located on the cell growing surfaces of the first passage 108. In some embodiments, the culture medium can deliver nutrients and oxygen to those cells as the fluid passes those cells in a laminar manner. This can facilitate healthy cell growth.

Once a desired amount of cell growth has been achieved, the cell growth facilitating fluid can be replaced with a second fluid adapted to cause the cells to dislodge from the cell growing surfaces. Exemplary second fluids contain, for example, dissociation fluids such as, proteolytic enzymes (e.g., Trypsin) and/or a metal atom and/or ion chelator (e.g. EDTA). After being added to the container 102, the second fluid flows from the upper portion 104 to the lower portion 106 over the cells in a substantially laminar manner. This causes the cells to dislodge from the cell growing surfaces of the first passage 108 and, eventually, exit the container via a drain connection (not shown) in the container 102.

According to the embodiment illustrated in FIG. 1, multiple plates 116a-l 16e are aligned in a stacked fashion between the upper portion 104 and the lower portion 106 of the container 102. Each plate 116a-l 16e includes surfaces that define at least one aperture 120 passing through it from an upper surface of the plate to a lower surface of the plate in a substantially vertical direction. Those surfaces form a portion of the first passage 108. A spacer 118a-l 18d (see FIG. 2) is positioned between each set of adjacent plates (e.g., 116a and 116b). Each spacer 118a- 1 ISd separates adjacent plates from each other, thereby defining a space between those adjacent plates (e.g., 116a and 116b). The space between each set of adjacent plates (e.g., 116a and 116b) also forms a portion of the first passage 108.

In the illustrated embodiment, the apertures 120 of adjacent plates 116a-l 16e that form portions of the same first passage 108 are not vertically aligned with each other. For example, the aperture 120 that passes through plate 116a to form a portion of the first passage 108 is not aligned with the aperture 120 that passes through adjacent plate 116b to form another portion of the first passage 108. The space between those adjacent plates 116a and 116b includes a substantially horizontal portion that facilitates fluid flow from the aperture 120 in plate 116a to the aperture 120 in plate 116b.

As illustrated, certain surfaces 11 Oa-11Oe of the first passage 108 are adapted for growing cells thereupon. Each of those cell growing surfaces 11 Oa-11Oe is disposed to facilitate fluid flow in a substantially horizontal direction. In some embodiments, the cell growing surfaces are, or include, a plastic material that has been tissue culture treated to facilitate cell growth thereupon. Different materials can be suitable for use as cell growing surfaces depending, for example, on the type of cells to be grown. As an example, polystyrene material, such as DOWs Styrone 666, can be suitable for use as cell growing surfaces in certain applications. Other suitable materials include, for example, polycarbonate, glass, plastics, and other materials. According to the illustrated embodiment, a pumping mechanism 114 is provided to motivate fluid flow from the lower portion 106 of the container 102 to the upper portion 104 of the container 102. As shown, the pumping mechanism 114 includes an air-lift pumping arrangement. Specifically, an air compressor 122 is coupled to the container 102. The air compressor 122 is adapted to draw air through a High Efficiency Particulate Air (HEPA) filter 124 and deliver compressed air via pipe 126 into a lower section of second passage 112. In the illustrated embodiment, the second passage 112 is vertically disposed in approximately the center of the container 102. The compressed air travels up the second passage 112 while drawing fluid up second passage 112 via an opening at the bottom of the second passage 112. In some instances, source air for the air compressor can come from inside an incubator via the HEPA filter 124. The air that is inside the container 102 and above the surface level 130 of fluid can diffuse into the fluid. The fluid delivers this air to cells growing upon surfaces HOa-llOe.

In various applications, other pumping mechanisms can be suitable to motivate fluid flow from the lower portion 106 of the container 102 to the upper portion 104 of the container 102. Examples of such other pumping mechanisms include displacement pumps, peristaltic pumps, and vacuum-driven pumps. In certain implementations, the choice of an appropriate pumping mechanism 122 can be influenced by a desire to maintain fluid sterility. In such instances, for example, it can be desirable to use a pumping mechanism 114 adapted to motivate fluid flow without physically contacting the fluid.

The illustrated embodiment also includes an optional heating element 127 positioned at a lower section and inside of the second passage 112. The heating element 127 is an electric resistance-type heating element. Other types of heating elements can be suitable for use in particular implementations. The heating element 127 is adapted to heat the fluid inside the second passage 112. That heated fluid tends to rise inside the second passage 112. The heating element 127 thereby facilitates fluid flow from the lower portion 106 to the upper portion 104 of the container 102. In an alternative embodiment, the heating element 127 can be coupled directly to a bottom surface 128 of the container 102. In that instance, the heating element 127 might be positioned adjacent the opening at the bottom of the second passage 112 so that the fluid heated thereby might tend to rise inside the second passage 112.

The heating element 127 also can be used to moderate the temperature of the fluid in order to provide a measure of control over the rate of cell growth on the cell growth surfaces 1 lOa-11Oe of the first passage 108, as would be readily understood by one of ordinary skill in the art.

Certain embodiments include surfaces inside the container that define a multiple first passages 108, each of which facilitates fluid flow from the upper portion 104 of the container 102 to the lower portion 106 of the container 102. In those instances, each of those first passages 108 might or might not be hydraulically isolated from the other first passages 108. Some implementations can include a container 102 that includes therein, surfaces that define multiple second passages 112, with each second passage 112 being adapted to facilitate fluid flow from the lower portion 106 of the container to the upper portion 104 of the container. Each second passage 112 can have an associated pumping mechanism 114 adapted to motivate fluid flow in that direction via the second passage 112.

The illustration shows a number of exemplary light sources 132 coupled to the container and adapted to transmit light into fluid inside the container 102. This transmitted light can influence a rate of cell growth for the cells on a cell growing surface 1 lOa-11Oe near an associated light source 132.

According to the illustrated implementation, an impedance sensor 134 is coupled to each cell growing surface 11 Oa-11Oe. The impedance sensor 134 can be adapted to provide information regarding the rate of cell growth and total number of cells upon each of those cell growing surfaces 11 Oa-11Oe. A controller (not illustrated) can be adapted to receive and process data from the illustrated impedance sensors 134. The controller can be adapted to adjust each of the light sources 132 independently, the heating element 127 and the flow rate/pressure associated with the pumping mechanism 114 in order to maximize cell growth. With reference to the implementation of FIG. 1, operation of the cell culture apparatus will be discussed. As mentioned above, the culture is initiated by adding cells suspended in a culture medium to the cell culture apparatus. The cells then settle onto, and adhere to the surfaces inside the apparatus that are adapted to facilitate cell growth thereupon. The pumping mechanism 114 is then activated. Fluid then flows in a substantially laminar manner from the upper portion 104 of the container 102 to the lower portion 106 of the container 102 along first passage 108. More specifically, fluid flows from the upper portion 104 of the container 102, through an aperture 120 that passes, in a substantially vertical direction, through plate 116a. That fluid then flows, in a substantially horizontal direction and in a substantially laminar manner, across cell growing surface 110b of plate 116b. That substantially horizontal portion of the first passage 108 is defined by plates 116a, 116b and spacer 118a. As the fluid flows across cell growing surface 110b, it delivers nutrients and/or air to the cells growing thereupon.

The fluid then flows through a substantially vertically disposed aperture 120 in plate 116b and into a second substantially horizontal portion of path 108 that is defined by plates 116b, 116c and spacer 118b. The fluid continues to flow along the path indicated by dashed line 108 until it reaches the lower portion 106 of container 102.

An air compressor 122 delivers filtered, compressed air into a lower portion of the second passage 112. The compressed air aerates fluid located inside the second passage 112 and helps to motivate the aerated fluid upward. At an upper end of the second passage, the aerated fluid spills out of the second passage 112 (indicated by arrows) and into the upper portion 104 of the container 102.

Gravity facilitates fluid flow in a laminar manner from the upper portion 104 of the container 102 to the lower portion 106 of the container 102 via the first passage 108. Also, as fluid is "pumped" up the second passage 112, a localized low pressure region is created in the lower portion 106 of the container 102 near the lower opening of the second passage 112. This also facilitates the flow of fluid through the first passage 108. As fluid travels through the first passage 108, it delivers nutrients and oxygen to cells that are growing on surfaces 11 Oa-11Oe. In some implementations, the container 102 is cylindrical and each plate 116a-

116e and each spacer 118a-l 18e is disk-shaped and sized to fit snugly within the cylindrical container 102. Referring to FIG. 2, an exploded view of disk-shaped plates 116a-l 16e and spacers 118a-l 18d that might be fit into a cylindrical container 102 is shown. Although the illustration shows an exploded view, when assembled, each plate 116a-l 16e is held in close proximity to its adjacent spacers 118a-118e.

Each plate 116a-l 16e is substantially flat and includes surfaces that define four apertures 120. Each aperture 120 extends from an outer peripheral surface 202 of its respective plate 116a-l 16e radially inward along a straight line toward a center point of the disk-shaped plate 116a-l 16e. Each aperture 120 terminates at a point that is proximate a center point of its respective plate 116a- 116e. As illustrated, the apertures 120 that pass through adjacent plates 116a-l 16e and define a portion of the same first passage 108 are angularly displaced from each other so that they are not vertically aligned with each other. For example, first passage 108 includes one aperture 120 that passes through plate 116a and another aperture 120 that passes through plate 116b. The aperture 120 that passes through plate 116b is angularly displaced from vertical alignment with the aperture 120 that passes through plate 116a by approximately 90 degrees.

Each spacer 118a-l 18d includes an outer rim 204 that defines a circular pattern and an inner rim 206 that defines a smaller circular pattern. Four spokes 208 extend between each outer rim 204 and each inner rim 206. Each spoke 208 extends from its associated outer rim 204 along a substantially straight line toward its associated inner rim 206. The substantially straight line is offset from an imaginary line that would diametrically dissect the circular pattern defined by the outer rim 204. As illustrated, each spacer 118a-l 18d includes four approximately wedge-shaped open sections that are defined by surfaces of the inner rim 206, the outer rim 204 and the four spokes 208. Put another way, the inner rim 206 and the outer rim 204 are each shaped as an annulus, with four spokes 208 connecting one annulus to the other, and with each spoke spaced apart by about 90°.

Each of the four apertures 120 that pass through plate 116a are associated with a corresponding first passage 108 in a container 102. Accordingly, the plate/spacer arrangement represented in the illustrated embodiment, if assembled, would include four separate first passages 108, each of which would extend from above plate 116a to below plate 116e. Each of those first passages 108 would run in a substantially spiral manner and each would be hydraulically isolated from other first passages 108.

When stacked together, the spokes 208 of each spacer HSa-I lSd are skewed a few degrees from associated apertures 120 in adjacent plates 116a-l 16e.

Accordingly, during operation, fluid flows downward through a first aperture 120 (e.g., aperture 120 of plate 116a) and hits an upper surface of the next lower plate in the stack (e.g., plate 116b). The fluid then flows around a portion of a circular path inside the disk-shaped container in a substantially horizontal direction between the lower plate (e.g., plate 116b) and the upper plate (e.g., plate 116a) . hi the illustrated embodiment, the portion of the circular path is approximately 90°. As the fluid flows along that portion of the circular path, it delivers nutrients and air to cells that are growing upon the upper surface of the lower plate (e.g., plate 116b). At about the end of the 90° portion, the fluid reaches a second aperture 120 that passes through the lower plate (e.g., plate 116b). The fluid flows downward through that aperture 120 and lands on an upper surface plate 116c. The fluid then flows in a substantially horizontal plane another 90° around the portion of the circular path defined between the now upper plate (e.g., 116b) and the lower plate (e.g., 116c). The fluid continues flowing through the rest of the plates in a similar manner and along a path that is indicated approximately by dashed line 108 in FIG. 2. Each plate 116a-116e and each spacer 118a-118d includes a surface that defines a hole 209 passing through approximately its respective center points. In one implementation, the holes 209 of adjacent plates and spacers align to form a second passage 112 when assembled. In such an implementation, a sealant can be included to prevent fluid flowing through the second passage from leaking out of the second passage. According to another embodiment, the holes 209 of adjacent plates and spacers align to create a space to accommodate a pipe that forms a second passage 112. Three alignment holes 210 are provided in each plate 116a-l 16e and each spacer 118a-l 18d to facilitate proper alignment of the stacked plates and spacers when assembly. FIG. 3 illustrates an alternative implementation of a cell culture apparatus 100.

A culture is initiated in the illustrated embodiment by adding cells suspended in a culture medium to the cell culture apparatus 100. The cells then settle onto, and adhere to those surfaces 11 Oa-11Oe inside the apparatus that are adapted to facilitate cell growth thereupon. The illustrated implementation includes a first passage 108 that follows a substantially zigzag pattern downward from the upper portion 104 to the lower portion 106. Adjacent plates (e.g., 116a and 116b) are arranged in a stacked fashion inside container 102 and separated from each other by spacers (not shown). Each plate 116a-l 16e includes at least one surface 1 lOa-11Oe for growing cells thereupon. Additionally, the second passage 112 that extends from the lower portion 106 of the container 102 includes a portion located outside the container 102. The

I l illustrated pumping mechanism 114 is an air-lift pump mat includes an air compressor 122 that draws air through a HEPA filter 124 and delivers that filtered air to a lower section of the second passage 112. The delivered air motivates fluid inside the second passage upward. That fluid spills out of an upper part of the second passage 112 and into the upper portion of the container 102.

FIG. 4 is a perspective view of a spiral-shaped element 404 having two surfaces 402a, 402b. The spiral-shaped element 404 can be positioned inside a container 102 to define first passages 108 between an upper portion 104 to a lower portion 106 of a container 102. When positioned inside a cylindrical container 102, a first passage 108 would be thereby defined on each side of the spiral-shaped element 404, each first passage 108 being associated with one or the other surfaces 402a, 402b. Each of those two first passages 108 would follow a substantially spiral and downward pattern. Cell growing surfaces 110a, 110b are provided on each of the first passages 108. In one implementation, the illustrated spiral-shaped element 404 would be snugly fit into a container 102, so that outermost edges of the surfaces 402s, 402b mate snugly against internal sides of the container 102. Such an arrangement might help ensure that fluid flows over the portions of the first passages 108 adapted to facilitate cell growth thereupon. In such an arrangement, a second passage 112 might pass outside the container 102 to connect a lower portion 106 of the container 102 to an upper portion 104 of that container 102. Alternatively, a second passage might pass upward through an axial center of the spiral-shaped element 404.

FIG. 5 illustrates a cutaway view of a particular implementation of a cell culture apparatus 100. The cell culture apparatus 100 includes a container 102 with an upper portion 104 and a lower portion 106 therein. Multiple plates 116 are arranged in a stacked manner inside the container 102. Adjacent plates 116 are separated from each other by spacers 118. A first passage is defined by surfaces of the plates 116 and separators 118 to facilitate laminar fluid communication from the upper portion 104 of the container 102 to the lower portion 106 of the container 102. A second passage is defined by surfaces on each plate 116 and spacer 118 that align to form a centrally disposed hole that passes from the lower portion 106 to the upper portion 102 through the stack of plates 116 and spacers 118.

In an exemplary embodiment, the container 102 of the cell culture apparatus has a diameter of approximately 5 inches and a height of approximately 9 inches. Such a container 102 might contain approximately 75 plates 116 therein.

Three ports 502, 504, 506 are provided at a lower surface of the container 102. Each port 502, 504, 506 is adapted to be press fit onto a complementary receptacle (not shown). According to one implementation, port 504 is a vent, port 506 is an air bubbling intake port and port 502 is a media in/out port. Mounting bases 508a, 508b, 508c, 508d also are provided at the lower surface of the container 102. Those mounting bases are adapted to be coupled to corresponding mounting features on a support structure (not shown). Such coupling can ensure proper alignment between the ports 502, 504, 506 of the container 102 and complementary receptacles on the support structure. Such coupling also can prevent unwanted lateral movement of the apparatus 100.

FIG. 6 illustrates a substantially closed system 600 for growing and harvesting cells. The illustrated system 600 includes a cell culture apparatus 100 and a cell harvesting unit 602. According to one embodiment, the cell harvesting unit 602 is a spin separation unit, which is a sealed unit with high efficacy (typically, greater than 97% of the cells are captured). In some embodiments, the same device is used to wash the cells, essentially flowing the fluid across the cells while they are in the spinner. An in-line cell counter 612 is provided between the cell culture apparatus 100 and the cell harvesting unit 602.

The system 600 includes a connection 604 for attaching a bag 606 containing a suspension of cells to be added to the culture apparatus.. The connection 604 is isolatable from the rest of the system 600 by valve 616a. A bag 606 of initiated cells is shown attached to the connection 604. The initiated cells are transferred to the cell culture apparatus 100 via line 608. Those cells are cultured in the cell culture device 100. After detachment from the surface using, for example, a proteolytic enzyme, cells can be removed from the cell culture device 100 via line 610. The in-line cell counter 612 is adapted to count the number of cells that are transferred from the cell culture apparatus 100 to the cell harvesting device 602. In one embodiment, the inline cell counter 612 is Laser Particle Counter PC 2000, manufactured by Spectrix, Inc., Redwood City, California. In one embodiment, the cell harvesting device is a Cytomate spin separator, manufactured by Nexell, Irvine California. Cells are transferred from the cell harvesting device 602 via line 614. A valve

616b is provided to selectively isolate a cell collection bag 618 from other parts of the system 600. The cell collection bag 618 can be filled, sealed and removed from the system 600 for use, for example, by a doctor.

FIG. 7 illustrates an exemplary support structure 700 and associated tubing adapted to support four cell culture apparatuses 100.

FIG. 8 illustrates an exemplary piping diagram for a single cell culture apparatus 100.

A number of implementations have been described. Nevertheless, it will be understood that various modifications can be made without departing from the spirit and scope of the invention. For example, there are a variety of possible paths between the upper and lower portions of the container through which the fluid might flow in a laminar manner. Additionally, the particular configuration of the second passage from the lower portion of the container to the upper portion of the container can vary. The particular placement of various ancillary devices, such as the light sources, impedance sensors and heating elements can vary or, indeed, be eliminated entirely. Various fluids can be suitable for use in a particular application. Various pumping mechanisms can be suitable for use in a particular application. In the upper portion of the container, air can diffuse into the fluid through a catalytic membrane. Air diffusion can be accomplished in other ways know in the art. The particular physical configuration of the plates and spacers can vary. For example, the plates can include a lesser or greater number of apertures than specifically disclosed herein. Additionally, the shape and size of the apertures can vary. Similarly, the number of, shape of and position of the spokes of the spacers can vary. Indeed, the plates and spacers can have completely different physical appearances than those described herein. Each plate can be integrally cast with an adjacent spacer. The size of the cell culture apparatus can vary, as can the number of plates and spacers used in a cell culture apparatus. Various connections can be provided at the cell culture apparatus. A variety of materials can be suitable to manufacture the container, the plates, the spacers, the cell growing surfaces, etc.

Accordingly other implementations are within the scope of the following claims.

Claims

What is claimed is:

1. A cell culture apparatus comprising: a container having an upper portion and a lower portion therein; first surfaces inside the container that define a first passage between the upper portion and the lower portion; second surfaces inside the container that define a second passage between the lower portion and the upper portion; and a mechanism adapted to motivate fluid flow from the lower portion to the upper portion through the second passage; wherein a portion of the first surfaces is adapted for growing cells thereupon.

2. The cell culture apparatus of claim 1 wherein the first passage comprises: a plurality of plates positioned in a stacked fashion between the upper portion and the lower portion; and at least one spacer positioned between adjacent plates to define a space therebetween.

3. The cell culture apparatus of claim 2 wherein each plate comprises at least one aperture therethrough and wherein the first passage comprises at least one of the apertures through each plate and at least a portion of the space between adjacent plates.

4. The cell culture apparatus of claim 3 wherein the plurality of plates is arranged in such a manner that the apertures of adjacent plates are not aligned with each other.

5. The cell culture apparatus of claim 3 wherein the container is cylindrical; wherein each plate is disk-shaped; and wherein each aperture is rectangular in shape, a longer side extending from a position proximate a center point of an associated plate radially outward along a straight line from the center point to an edge of the associated plate.

6. The cell culture apparatus of claim 5 wherein each plate includes a plurality of apertures extending outward along respective straight lines each of which is positioned to extend from the center point in a direction that is angularly displaced from a straight line associated with an adjacent aperture on the same plate.

7. The cell culture apparatus of claim 5 wherein the space between adjacent plates permits fluid communication between a first aperture of an upper plate and a second aperture of a lower adjacent plate.

8. The cell culture apparatus of claim 7 wherein the portion of the first surfaces for growing cells thereupon comprises at least a portion of the space between adjacent plates.

9. The cell culture apparatus of claim 7 wherein the first aperture and the second aperture are not vertically aligned with each other.

10. The cell culture apparatus of claim 2 further comprising a support element positioned inside the container and having support shelves to support the plurality of plates and the at least one spacer; wherein the second passage is positioned inside the support element.

11. The cell culture apparatus of claim 1 wherein the mechanism adapted to motivate fluid flow is an air lift pump coupled to an entry port of the second passage located proximate the lower portion of the container.

12. The cell culture apparatus of claim 1 wherein the mechanism adapted to motivate fluid flow is a heating element adapted to heat fluid located proximate an entry port of the second passage located proximate the lower portion of the container.

13. The cell culture apparatus of claim 1 wherein the first passage is arranged in a manner to accommodate laminar flow of fluid therethrough.

14. The cell culture apparatus of claim 1 wherein the second passage is positioned at least partially outside the container.

15. The cell culture apparatus of claim 1 further comprising an impedance sensor coupled to the surface for growing cells, wherein the impedance sensor is adapted to measure impedance associated with the surface for growing cells.

16. The cell culture apparatus of claim 1 further comprising: a fluid inside the container; a heating element coupled to the container to heat the fluid inside the container; and a control unit adapted to modulate a rate of cell growth in the container by modulating a temperature of the fluid inside the container with the heating element.

17. The cell culture apparatus of claim 1 further comprising a fluid inside the container; a light source coupled to the container and adapted to transmit light into the fluid inside the container; and a controller adapted to control operation of the light source to modulate a rate of cell growth inside the container.

18. A method of growing cells comprising: adding cells to the apparatus of claim 1; and incubating the apparatus.

19. The method of claim 18 wherein the cells are fibroblasts.

20. The method of claim 18 wherein the cells are human cells.