JPH05501953A - Continuous high-density cell culture system - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Continuous high-density cell culture system TECHNICAL FIELD The present invention relates to a method for culturing cells and an apparatus for the same.

Traditionally, cells are mounted in glass or plastic roller containers and flasks. It was growing. This method is suitable for high-density cell culture or continuous cell culture. It is unsuitable and requires large amounts of media and space. Furthermore, this method is labor intensive. It is true.

To achieve high-density growth conditions, the stacked plate structure Various attempts have been made to ensure that the surface of the plate provides a large surface area for cell seeding and growth. is being done. Despite these various attempts, all prior art cell culture devices Requires excessive amounts of media; continuous flow of nutrients to all cell growth surfaces It requires labor-intensive monitoring and protection of the proliferating cells. Some disadvantages such as being difficult to operate and not being able to function continuously One objective of the present invention is to increase the density of cells relative to the amount of enrichment medium maintained in the system. An object of the present invention is to provide a cell culture device that can be used for cell culture at the same time.

Another object of the invention is to provide a cell with a plurality of growth chambers that evenly accepts the medium flow. Another object of the present invention is to provide a cell culture assembly.

Another object of the invention is to automatically and continuously add the nutrient medium and form it by the cells. A cell culture medium that allows removal of spent media, including produced products and waste. The goal is to provide a nourishing assembly.

Another object of the invention is to continuously monitor the environmental conditions of the cells and detect deviations from the desired state. Provide cell culture assemblies and systems that automatically correct or issue an alarm when Is Rukoto.

Another object of the present invention is to promote a desirable constant cell culture environment for the cells and to Ideal for cell products such as biochemicals, vaccines, viruses and drugs. The objective is to provide a continuous flow system that facilitates efficient yield and collection capabilities.

The present invention defines an extremely large surface area that allows for high density cell growth within a small volume. The present invention provides a continuous cell culture device with a row of growth chambers.

This device allows flow in a predetermined direction through the growth chamber, and the flow Structures and configurations that allow continuous access to all growth surfaces within each chamber. shall be established. Control fluid flow and distribution to each of the growth chambers to each chamber Ensuring sufficient flow within each chamber by providing fluid restriction ports that function can be realized.

The cell culture device is defined by spaces between a plurality of stacked plates. It is desirable to have a single row of cell growth chambers. The inlet conduit is a fluid restriction port provides a source of nutrient medium for the manifold, which is in fluid communication with the cell growth chamber through the do. The total cross-sectional area of the narrowest area of the fluid restriction port is less than the cross-sectional area of the inlet conduit. This reduces the pressure drop at each fluid restriction port and the flow of fluid through each growth chamber. Ensure adequate flow.

Fluid throttling ports and growth chambers allow fluid media to reach all growth surfaces with minimal turbulence. be structured and configured to facilitate distributed and continuous flow in a predetermined direction; This is desirable. This is the opposite side of the box to the proliferation chamber, which is a roughly square box. This is achieved by providing an inlet throttle port and an outlet throttle port at the corner. I can do that. These fluid restriction ports can have a narrow intermediate area and The inner portion may include a curved surface to facilitate fluid distribution and turbulence-free flow. I can do that. Ribs are provided between opposing surfaces of the plates to provide structural support for fluid flow, Furthermore, the direction of flow can be determined.

Thus, the present invention provides a closed sterilization system for cell culture, allowing humans to cells and their products and protect cells from contamination by the external environment. It is intended to prevent The proliferative state automatically senses and flourishes in response to the sensed state. Nutrients can be added and cell products removed. The device is economical to manufacture It is also possible to mass produce.

Brief description of the drawing Figure 1 is a perspective view from the upper right of the cell culture container of the present invention, and Figure 2 is taken along the line in Figure 1. A perspective cross-sectional view along a-a, FIG. 3 forming the cell culture vessel of FIG. A perspective view from above of a single culture plate used for Figure 4 is an enlarged view of the corner of the plate in Figure 3, and Figure 5 is an enlarged view of the wrapped culture tube. The present invention has the proximal side of the device casing cut away to show the surfaces and edges of the chamber. A perspective view of a cylindrical embodiment of a cell culture vessel, FIG. 6, shows the spiral growth chamber of FIG. An end cross-sectional view of the chamber, FIG. 7, shows cells of the invention in a continuous cell culture assembly. A schematic diagram of a culture vessel, FIG. 8, shows the plate used to form the culture vessel of FIG. plate assembly with gaskets defining the height and shape of the space between the plates; FIG. 2 is an exploded front view of one embodiment of the body.

Brief description of the drawing A preferred embodiment cell culture device includes an array of growth chambers enclosed within a container. It is growing. The growth chamber provides a large surface area for cells to grow.

In the external view (FIG. 1), the container 10 has a square shaped top 12 and a bottom 13. It is a generally square box with vertical molded side walls 14 sealed along the edges and a growth chamber. Provides a fluid-tight mechanism for housing the bar. An inlet conduit 16 and an outlet conduit 18 are connected to the top 1 Extending from diagonally opposite corners of 2. The inlet conduit 16 is connected to the inner space of the container 10. The outlet conduit 18 provides fluid access to the interior space of the container 10. Provide a fluid outlet for both. The outlet conduit 18 is not from the top as shown. , may extend through the base 13.

Referring to FIG. 2, the inlet conduit 16 opens the interior space of the container 10 at a corner. axially aligned and in fluid communication with an inlet manifold 20 extending laterally therethrough. There is. The inlet throttle port 22 is connected to a plurality of ports located within the manifold 20 and the vessel 10. in fluid communication with a cell growth chamber 24, extending upwardly and downwardly from the cell growth chamber 24; The degree is defined by a row of stacked plates 26. Outlet manifold 28 comes out. located in the knee of the container opposite the inlet manifold 20 in fluid communication with the inlet conduit 18; I'm being kicked. Each of the growth chambers 24 has an outlet through an outlet fluid restriction port 23. It is in fluid communication with the mouth manifold 28. Inlet and outlet manifolds in preferred embodiments The hold mechanisms are mirror images of each other and, therefore, their flow characteristics depend on whether the fluid medium is at the inlet end or It is the same whether the flow is from the outlet end to the inlet end or vice versa. be.

The relative dimensions of the throttle port 22.23 and conduit 16.18 are an important part of the invention. form. Each chamber receives continuous and controlled fluid medium flow. To ensure a pressure drop at the flow restriction ports that control the flow within each chamber, It is necessary to cause it to occur. To achieve this, an aperture point that controls the flow is required. The total cross-sectional area of the narrowest part of these throttle ports is equal to the cross-sectional area of the inlet conduit. or preferably less than or equal to . These total cross-sectional areas are also , preferably equal to or less than the cross-sectional area of the exit conduit. These conditions In some cases, no particular growth chamber placement location is particularly desirable; The expansion chamber primarily has a special fluid restriction port that controls the flow within that chamber. Accepts fluid media based on dimensions. These fluid restriction ports have uniform dimensions and each growth chamber can receive an equal flow rate of fluid medium. desirable.

In the illustrated embodiment, flow restriction ports are provided at both the inlet and outlet ends of the growth chamber. A route is provided. The smaller of these inlet and outlet restriction ports Control flow through a particular growth chamber, whether at the oral end or the eight-end . In this way, the smaller total cross-section of the inlet and outlet restrictor ports for each chamber The area must be smaller than the cross-sectional area of the conduit. A pair of inlet and outlet throttle ports If the dimensions are uniform and the cross-sectional area of the narrowest part is equal, then the total cross-sectional area mentioned above is When calculating, include only one cross-sectional area in the calculation.

Plates 28 are stacked to form a growth chamber. these plates It is desirable that they be separated by 1 mm or more. These plates are closer together However, if the spacing is less than 1 mm, air bubbles may occur between the plates. is incorporated and tends to disturb the uniform flow of the medium. The plate surface is rough wavy, spiral or other irregular shape to increase its surface area; , it is possible to increase the number of cells that can be grown on a given plate. make it irregular In this case, the 1 mm spacing between the plates is the distance between the facing vertices of their irregular surfaces. be the spacing dimension. In addition, the plate surface may promote cell proliferation. Sea urchins can be surface treated in a variety of different ways. Typical processing involves Includes Bonyl group treatment, collagen treatment, fibroma treatment or feeder cell layer.

Plates according to preferred embodiments are manufactured by F.F., Bartlesville, Oklahoma 74004. Phillips Chemical Company C Polystyrene and butadiene resins and blocks sold by Molded with black copolymer. Suitable plate materials include styrenic or polymeric materials. Contains materials such as chillpentene. However, the plate is sufficiently robust and Virtually any material that is non-toxic, biocompatible, and otherwise suitable for culturing cells. It is possible to form it with the material of.

The preferred embodiment plate has a novel structure that facilitates flow distribution and manufacturing. ing. Each of the plates 26 has an upper surface 30 in contact with an outer peripheral wall 32 (a third Figure 3). The inwardly facing surfaces 34 of the walls 32, together with the top surface 30 of the plate, form the growth chamber 2. 4 side walls and a floor. When plates 26 are stacked in the assembled device , the apical edge 35 of the wall 32 engages and seals with the lower surface 36 of the adjacent plate, causing the growth It defines a chamber, the ceiling portion of which is defined by the lower surface 36 of the adjacent plate. is defined. The top end of the wall 32 to facilitate mating placement of the stacked plates. Edge 35 is formed with a groove 33 that receives an engagement ridge 37 on the bottom of the adjacent plate. will be accomplished. These engaging ridges 37 and grooves 33 are aligned with the stacked plates. do. The four corner portions 38 and 40 of each plate 26 are equal to the height of the outer peripheral wall 32. Fill the mixture until it reaches the desired thickness. The filled corners are attached to the assembled structure. Provides support. The filled corner is, as explained in more detail below, Further reducing fluid turbulence and "dead spots".

Each plate 26 has holes in diagonally opposed filling corners 40. Assemble In the upright position, the holes 42, 44 in the stacked plates 26 are axially aligned. and form an inlet manifold 20 and an outlet manifold 28, respectively. aisle 48 from each hole 42, 44 through the filled corner 40 to the side wall of each plate 26. and extending into the interior space defined by the floor. In the assembled state The passageway provides fluid access between the manifold 20.28 and the growth chamber 24. A fluid restriction port 22.23 is formed. Each of these passages is connected to a plate 26 and the floor defined by a part of the upper surface 30 of the corner part 40. The side wall 49 has a height equal to that of the outer horn 32. Opposing Stack the flat bottoms of the plates 26 and seal the open tops of the passageways 48. Sometimes a fluid restriction port 22.23 is formed.

The particular shape of the fluid restriction port in the preferred embodiment is such that the It has a small inner part. In this case, the diameter of this flow restriction port 22 is It increases continuously and substantially in the direction towards the chamber 24. With such an arrangement, The medium entering the growth chamber spreads outward and forms a radial pattern within the space of the growth chamber. Uniform distribution of nutrients and fluid flow over the entire growth surface Promote this. In addition, this configuration particularly reduces turbulence near the throttle boat 22. It is something to do. In the illustrated embodiment, the diameter of the flow restriction port increases linearly. are doing. However, a port formed as a second or third venturi continuously and progressively change the flow rate, thereby further promoting turbulence-free flow. Ru. A flow pattern in a predetermined direction without turbulence is achieved by filling the corner 38 and This is further facilitated by the guide ribs 46. If this corner is not filled , providing a wall perpendicular to the flow direction, such a wall resulting in a growth chamber containing: Turbulent flow and “dead spots” where new medium flow is not supplied continuously and in a given direction. ”, which can lead to cell death. The filled corners are A curved surface is provided which acts to continuously guide the flow toward the exit port. rib 4 6 serves two functions. These ribs can be used to drain liquid from the device using vacuum force. Provides structural support for each plate while maintaining proper spacing. , and positioned so as to continuously direct the flow toward the exit boat. I can do it.

To assemble the device, the plates are placed with their outer peripheral walls and corner portions facing each other. stacked and secured to each other in such a way that they can be sealed to the bottom of the plates; It takes. Currently, each of these plates is fused to the adjacent plate using ultrasonic welding. are in contact with each other. However, the final product has sufficient strength and is harmless to the cultured cells. As long as any fabrication such as solvent welding, RF welding, botting, glue or gaskets construction method can be adopted.

In operation, the device is first seeded with cells. Next, apply fluid to the seeded cells as follows: Supply medium. The fluid medium is conducted into the inlet manifold 20 via the inlet conduit 16. Enter. The fluid medium is then directed through the various flow restriction ports 22 to the associated growth channels. Enter chamber 24. Because of the flow restriction ports and the overall structure of the growth chamber, The flow is continuously and completely distributed over all surfaces of the growth chamber 24, and the fluid medium is , always moves in the direction of the outlet flow restriction port 23. Next, the medium is the outlet flow restrictor. It enters the outlet manifold 28 through port 23 and exits the device through outlet conduit 18. come out.

Another embodiment of the device holds a longitudinally oriented line of culture chambers. It has a cylindrical casing. As shown in FIG. 6 envelops the cylindrical culture chamber 57 and is sealed to its outer surface. Keishin The plate 56 has two plates at each end, an inlet end plate 58 and an outlet end plate 59. are sealed into circular end plates, each of which is connected to a culture chamber 57. is slightly spaced from both ends. The inlet end plate 58 of the culture chamber 57 and The spacing between the mouth end defines an inlet manifold 60. Outlet of culture chamber 57 The spacing between end plate 59 and the outlet end defines an outlet manifold 62. fluid is fluidly attached to and from the inlet end plate 58. It enters the inlet manifold 60 via an axially extending inlet conduit 64 . The fluid comes out Fluid flowably attached to and axially extending from the mouth end plate 59 Exiting from the outlet manifold 62 via an outlet conduit 66 extending to.

The longitudinally extending growth channel of the culture chamber 57 has a plurality of longitudinally oriented growth channels. Formed by rolling a rectangular flexible sheet 72 formed with several protruding ridges 74. be done. When the sheet is rolled from the side parallel to the protruding ridge 74, the sheet 7 2 has a spiral cylindrical shape (FIG. 6). The protruding ridge 74 is a rolled sheet. together with the overlapping portions of the ridges 72 define a longitudinal channel row 76; 9 defines the height of each channel 76. The open end of channel 76 may be connected to a stopper or Restriction port 2 capped and restricts fluid access to each channel 76 Provide 2.

The operation of this embodiment is similar to that described above. First, the device is inoculated with cells, The cells are allowed to adhere to the surface of channel 76. Next, the fluid medium is introduced into the inlet. It is introduced into the inlet manifold 60 via tube 64. Next, the fluid exposure area is inlet throttle Proceeding from the manifold through port 22 into a longitudinally extending growth chamber 76. nothing. The fluid field runs continuously along the length of the growth channel 76 and reaches the exit aperture point. (not shown) to the device via outlet manifold 62 and outlet conduit 66. come out.

According to one feature of the invention, the cell growth device of the invention is shown schematically in FIG. as part of assembly 80. Assembly 80 is capable of continuous operation. The structure and composition shall be as follows. Assembly 80 connects culture vessel 10 of the present invention to a fluid reservoir. It is a closed loop system that connects to the server 82. Reservoir 82 outlet boat 84 is connected to the inlet port 86 of the culture vessel 10 via a fluid supply conduit 88 .

The outlet end 90 of the culture vessel 10 connects to the inlet port of the reservoir 82 via a fluid return conduit 94. In fluid communication with port 92. A pump 96 is disposed along fluid supply conduit 88. The fluid flow is continuously delivered from the reservoir through the culture vessel 10. nutritional supply A tube 98 fluidly connects to the fluid supply conduit 88 between the pump 96 and the container 10. , thereby allowing nutrients to be provided to the fluid supply conduit 88 . nutrition A pump 100 is provided for pumping fluid into fluid supply conduit 88 . Also, reservoir 8 2 includes a suction pump 104 that preferably continuously removes fluid downstream of the culture vessel. A product suction conduit 102 is provided which is connected to. Finally, the assembly includes a fluid Probes 106 are provided in supply conduit 88 and fluid aspiration conduit 94. these pros The probe 106 provides a means for sensing field conditions. These probes control While the control device 108 can be connected to the device 1if108, the control device 108 can be connected to various ports. It connects to a pump and allows the introduction of nutrients and absorption of products from the reservoir based on field conditions. control the pull. Optionally, an oxygen exchanger that continuously re-oxygenates the culture field It is also possible to provide exchange means 110. Nutrient feed pump 100 and product suction The pump 104 is operated continuously and at a constant speed so that the resupply field is continuously introduced. The product is continuously drawn from the system and its delivery rate is controlled by control means. It is desirable to have it set.

As mentioned above, the flow restriction ports need not be uniformly sized. each chamber If it is desired to have uniform flow through, the flow restriction ports are generally equal It is necessary to make the diameter. However, those skilled in the art will only know the dimensions of the flow restriction port. It is understood that the specific dimensions of the growth chamber also determine the flow rate. Good morning. In this way, the relative dimensions of the flow restriction port and the growth chamber are both While maintaining a constant flow rate, it is also possible to vary it.

In the preferred embodiment, only one pair of aperture ports per growth chamber is described. did. Of course, it is possible to provide multiple flow restriction means on the inlet or outlet side. . The inventor has now discovered that flow restriction ports are easier to manufacture, less turbulent, and typically One point is that it can provide results that alleviate the lack of cell proliferation seen in areas very close to It is considered desirable to provide inlet and outlet restriction ports.

Furthermore, it is possible for a person skilled in the art to provide a flow restriction port only on the inlet side. will be understood. Traditional methods of removing cells from culture vessels require a narrow outlet that is blocked. Containers with such constructions tend to create cell clumps that can cause cells to Desirable for applications that require removal from the container. Also, exit the aperture port. It is also possible to provide it only on the mouth side. However, currently, the flow restriction port is It is preferable to install it on the outlet side.

The total cross-sectional area of the flow restriction ports shall be less than or equal to the cross-sectional area of the outlet conduit. It is even more desirable that Otherwise, the outlet conduit will be subject to back pressure, resulting in will be preferentially carried out through one or more culture chambers.

In another embodiment, a cell culture vessel similar to that shown in FIGS. A side wall of the growth chamber (FIG. 8) and a gasket sandwiching the aperture port. It can be formed from a series of flat plates. In this example, the plate 126 has generally flat top and bottom surfaces. Cross holes 128 are located in each plate. located at each of a pair of opposing corners. The gasket 130 is a plate It has an outer periphery with a size and shape corresponding to the outer periphery of 126. gasket 130 Interior surface 132 defines a hollow space. A pair of plates 126 are gaskets 13 0, the opposing surfaces of plate 126 extend above and below the growth chamber. The inner wall 132 of the gasket forms the sidewalls of the growth chamber. Ru.

The corners 134 of the gasket are filled. Each of the pair of opposing corner portions is penetrating. The gasket holes 136 extend through the plate. The hole 128 is sized and shaped to correspond to the hole 128 in the opposite corner of the hole 126 . aisle 138 passes from the gasket hole 136 through the corner to the inward wall of the gasket 130. Extending inwardly to an interior space defined by 132. A pair of plates 12 6 clamps the gasket 130, the hole 128 of the plate 126 and the gasket The cut holes 136 align to form a manifold. Passageway 138 is a The rate is occluded at the upper and lower extensions to form a restriction port.

Such stacked plate and gasket assemblies are subject to compressive forces such as clamps. They can be held together by This particular embodiment of the invention is directed to the growth of cells. This has the advantage that the plates can then be separated from each other. This allows the cell A single layer or “skin” of Additionally, cells on the plate surface can be easily accessed.

The cell culture assembly used in this example has approximately the same structure as that shown in FIG. (excluding probe and oxygen exchange means).

The cell culture container used in this assembly had the following characteristics. 10 pieces The plates were stacked and secured together by ultrasonic welding. The bottom plate of the container I tried to form the bottom part. Inlet and outlet conduit fittings are provided on the top plate with points The plate was sealed to the top plate so that the top growth chamber was formed.

This top plate has a potting structure on all four sides of the stacked plates. The shape clamped and secured it in place. Each plate has a thickness of approximately 1. 5 mm, each plate having walls approximately 1 mm high with filled corners. I did it like that. When stacked, the plates are approximately 1mm high and 22mm long and wide. A growth chamber was formed. The narrowest part of the aperture port is 1mm in height x 2 in width. mm, defining a cross-sectional area of approximately 2 mm2. The cylindrical inlet conduit has a diameter of approximately 5 m. m and defined a cross-sectional area of approximately 49 mm2. Thus, the inlet conduit Cross-sectional area (49 mm is the total cross-sectional area of the smallest part of the aperture port (20 mm)) The result was higher than that.

Cells used in this example were obtained from American Thai, Rockville, Murrayland. American Type Cu1ture Co11ection) as ATCC No. CCL-81. (VEIiO) cells. These Vero cells are available in roller bottles (Massachusetts). Costar Corp., Cambridge, Ts. cells at a density of 5. Ox 10'. And so. The cells are then treated with tryptonite under aseptic conditions to remove the cells from the bottle surface. cells were removed and then placed in 15 ml of MEM, Grand Island, NY. Gjbco) and 5ml FBS (St. Louis, Missouri) Fetal Bovine Serum rum, Sigma)).

The assembly was filled with 1.250 ml of fluid field (MEM in 5% FBS); The vessel is then injected with cells through a sterile septum placed within the fluid supply conduit on the same day. Load the cells by loading the fluid, while the fluid is introduced into the container by a pump. Ta. This loading requires that the container be placed in its normal working position (inlet conduit at the bottom and outlet conduit is at the top and its sides are vertically raised) . In this position, all bubbles entering the container will rise and exit the container. fluid cells Operate the pump continuously during gradual injection into the open stream, and the pump and cells injection is approximately the time required to move the field from the injection point to the outlet port of the vessel. It was stopped when 90% had passed. This cell loading method is performed evenly during injection. dilute to uniformly treat cells in all growth chambers of the vessel. next , the container is left in contact with its bottom surfaces for 4 hours, allowing the cells to adhere to that surface. I did it like that.

Next, position the container so that its top surface is in a horizontal position and allow the cells to attach to that surface. The loading method described above was repeated, except that it was allowed to

After seeding the container with cells and allowing the cells to attach, pump 88 is started; It was operated continuously at a speed of 75 ml/min during 24 hours. Start on the second day and then By operating during a 24-hour period, additional fluid exposure is introduced into the system; Its total volume was 1.750ml. Furthermore, on the second day, the pump speed was 75 m. The speed was increased from 1/min to 150 mI/min. Next, on the third day, the pump was 300m1 The speed was increased to a speed of /min, and it was operated continuously at that speed until the 5th day. of the pump On the fifth day of operation, 5% carbon dioxide gas was circulated over the fluid medium surface in the reservoir ( Approximately 25ml/min). On the fifth day of growth, the containers were removed from the assembly and rinsed with salt water. . Thereafter, glutaraldehyde (in saline) was introduced into the container to fix the cells.

Cells were then colored with flight dye. After coloring, colored cells The container was cut open for inspection. The plate surface is a uniform layer of cells. It was completely covered.

Glucose consumption was monitored during the 5 day growth period. The amount of glucose consumed is It was as follows.

During the 5-day growth period, the rate increases in response to changes in glucose consumption and new A large open field was introduced through a nutrient supply conduit. Reservoir the corresponding amount of used field was removed via the overflow conduit within the tank. The capacity of the system is I, 750m. 1, this increment of 1 or more resulted in an overflow. Therefore, Once the system is completely filled, by adding fresh fluid flow, This resulted in a corresponding equal amount of spent media being removed from the fluid reservoir.

Glucose is monitored by inserting a syringe into the septum and removing a small fluid sample. I looked at it. The glucose level of this specimen is then tested at a location remote from the cell culture assembly. I tried it. Additionally, as described herein, the assembly may be continuously supplied with glucose, etc. It is also possible to provide on-line probes for monitoring.

It is understood that various modifications and variations of the embodiments described above are possible within the scope of the present invention. should be understood. All matter contained in the above description or shown in the accompanying drawings is provided by way of example only. It should not be interpreted as having a limited meaning.

FIG, I FIG. 2 FIG.3 FIG.4 FIG. 5 FIG. 6 international search report

Claims (16)

[Claims] 1. As a tissue culture device, A plurality of cell growth chambers, the inner surfaces of which are suitable for cell growth. a cell growth chamber; an inlet conduit providing a source of fluid to the chamber and discharging fluid from the chamber; an exit conduit for discharging the flow through at least one of said conduits and each of said growth chambers; and a fluid restricting means for communicating with the body, the total cross section of the narrowest part of the fluid restricting means characterized in that the product is equal to or less than the cross-sectional area of the inlet conduit. Tissue culture equipment. 2. The tissue culture device according to claim 1, wherein the cell growth chamber is multiple. Tissue culture characterized in that it is defined by a spacing between a number of stacked plates. Device. 3. The tissue culture device according to claim 2, wherein the fluid restriction port and the front The blade moves the fluid continuously and in a predetermined range from the inlet end to the outlet end of the chamber. A tissue characterized by having a structure and composition that allows it to flow in any direction without turbulence. Culture device. 4. The tissue culture device according to claim 2, wherein the plate comprises at least A tissue culture device characterized in that they are separated by 1 mm. 5. The tissue culture device according to claim 2, wherein the plate is approximately square. and the fluid restriction means includes fluid restriction ports in a pair of opposing corners of the plate. A tissue culture device comprising: 6. The tissue culture apparatus according to claim 2 is used to separate the plate. A tissue culture device further comprising support ribs. 7. Assemblies stacked together with similar plates to define multiple tissue culture chambers In a woven culture plate, A molded rectangular platform having generally flat top and bottom surfaces. a platform, at least one of the surfaces including a peripheral ridge; The seat has a pair of diagonally opposing corner portions, and the corner portion is at the height of the outer ridge. filled up to Furthermore, a hole provided in each of the filled corners, the hole being formed by the platform. a hole having an axis perpendicular to a plane defined by the hole; and a second passage communicating with the inner space defined by the foam. A tissue culture plate featuring: 8. The tissue culture plate according to claim 7, wherein the plate A tissue culture plate further comprising a support rib integral with the top surface of the plate. 9. providing a plurality of growth chambers and fluid access to the growth chambers; an inlet port; a narrow intermediate portion; and a tissue culture device comprising: further comprising an inlet port having a diameter increasing in the direction of the chamber; The fluid flowing into the growth chamber is distributed in a wide angle radial pattern. A tissue culture device characterized by: 10. The tissue culture device according to claim 2, wherein the cell growth chamber is defined by the spacing between multiple stacked plates separated by gaskets A tissue culture device characterized in that: 11. The tissue culture device according to claim 10, wherein the gasket and the plastic the plates may form a stacked row, and the plates may be separated from the stacked row A tissue culture device characterized by having the structure and configuration as follows. 12. The tissue culture apparatus according to claim 10, wherein the plate and the gas further comprising means for releasably securing the packets in stacked rows. Tissue culture equipment. 13. The tissue culture device of claim 1, wherein the inlet conduit and the plurality of further comprising an inlet manifold disposed between the cell growth chamber and the fluid restrictor. means are disposed between the inlet manifold and each of the cell growth chambers. A tissue culture device characterized by: 14. The tissue culture device according to claim 13, wherein the fluid restriction port and and said plate conveys fluid continuously and locally from an inlet end to an outlet end of said chamber. It is characterized by a structure and configuration that allows the flow to flow in a certain direction without turbulence. Tissue culture equipment. 15. The tissue culture device according to claim 13, wherein the narrowing means has a narrow and a diameter increasing in the direction of the growth chamber, thereby making it possible to The fluid flowing into the proliferation chamber through the inlet port has a wide angle radial pattern. A tissue culture device characterized in that the tissue culture is dispersed in a container. 16. The set according to any one of claims 1, 13, 14, or 15. a tissue culture device, an outlet disposed between the outlet conduit and the cell growth chamber; further comprising an outlet manifold, the throttling means connecting the outlet manifold and the growth chamber. What is claimed is: 1. A tissue culture device characterized in that the tissue culture device is disposed between a tissue culture device and a tissue culture device.