JPH01196289A - Vessel for cell culture - Google Patents

Abstract

PURPOSE:To provide the title vessel aimed at improving the collection and recovery of large quantities of cells subjected to culture, designed to install therein a cell culture tank equipped with a porous support, a culture solution discharge tank, a culture solution feed port, a cell recovery port and so on. CONSTITUTION:Firstly, a closed vessel 2 is equipped with a cell support (a porous slab) 1, a culture tank 9, and a culture solution discharge tank 10 provided with a culture solution discharge port 4 and a backwashing liquid feed port 5. Secondly, a culture solution 8 is fed, via a feed port 3, into the tank 9 to make a culture of cells 7. Thirdly, the waste materials and/or biologically active substances or the like produced as a result of the culture are penetrated through the porous slab 1 and discharged out of the system via the port 4. Finally, sufficiently grown cells 7 are easily separated from the porous slab 1 by feeding backwashing liquid through the port 5 and recovered via a cell recovery port 6, thus keeping the interior of the culture tank 9 under the stabilized environment for culture.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to a cell culture vessel for culturing cells in large quantities and for easily harvesting and recovering the cultured cells.

[Prior Art] Conventional cell culture containers include (1) culture bottles, roller bottles, (2) Sterilin type, (3) multilayer flat plates, and (4)
) multi-stage single layer, (5) hollow fiber, (6) spinner bottle, (7) beads, etc.

(1) is B 1otechno1. Bioeng,
, 10, p589 (1(2) is Exp, Ce1l
Res, 71, p293 (197(3) is B 1
otechno1. Bioeng, , 10, p.
601 (1(4) is protein nucleic acid enzyme, 21.p322
(1976) (5) 5science, 178. p65
(1972) (6) J, Im+l1uno1. ．． 79
．． p428 (1957) (7) is So+*atic C
e1l Genetics, 3, pi 49 (1
977) and others.

Problems to be Solved by the Invention Conventionally, this type of culture vessel has been used for the purpose of proliferating cells in order to obtain substances secreted by the cells.

In other words, it was designed to take out cells that produce substances from within a living body, grow them in an artificial environment, that is, in a culture container, increase their number, and collect the substances secreted into the culture medium along with the culture medium. . However, the secreted substance was soluble in the culture medium, and the secreted substance could be recovered by collecting the culture medium, but the target 5aua itself was not soluble in the culture medium, and it was not contained in the culture vessel. It is attached or deposited. In addition, since adhesion and deposition are essential conditions for cell proliferation, in order to efficiently collect and recover cells, remove cells from the cell support in the culture container when cell proliferation has progressed. It was necessary to separate the cells and collect and recover the cells floating in the solution.

[Example] Fig. 1 shows an example of the present invention, in which a closed container 2 is connected to a culture tank 9 in which cells are cultured using a cell support (porous membrane) 1, and is rich in waste products that have passed through the porous membrane. A culture solution discharge tank IO for discharging the culture solution is divided into a plurality of tanks. A culture solution supply port 3 and a cell collection port 6 are connected to the culture tank 9.
The culture solution discharge tank IO has a culture solution outlet 4 and a backwash solution supply port 5.
is in contact.

The structure shown in Figure 1 allows cell culture to continue in a stable state, and is effective in easily separating sufficiently proliferated cells from the cell support by backwashing them. It is possible to collect and recover cultured cells with high efficiency.

A constant flow of culture solution is always added through the culture solution supply port 3, and by adding the sterilized culture solution, the inside of the closed container 2 is always kept in a clean state. Waste products, physiologically active substances, etc. secreted by the cells can be eliminated by permeating the cell support l, which is a porous membrane, together with the culture solution, as shown in FIGS. 2 and 3. Therefore, the inside of the culture tank 9 works effectively to maintain a stable culture environment at all times (.Furthermore, with this configuration, the cell supports can be backwashed from the culture solution discharge tank 10 side, so that the cell growth has sufficiently progressed. It works effectively for separating cultured cells from the support l.Furthermore, it is useful for sterilizing the backwash solution and reducing the pore size of the cell support l by aseptically suspending the cells in the solution and suspending them. It is possible to collect and collect cells together with the liquid.As shown in Fig. 4, there is only one port that connects to the culture tank 9 of the closed container 2, and the culture medium supply port 3 and the cell collection port 6.
It is also possible to use a configuration in which the switch can be switched using a valve or the like.

Further, as shown in FIG. 5, a structure may be adopted in which there is one port communicating with the culture solution discharge tank 10, which serves both as the culture solution discharge port and the backwash solution supply port, and can be switched by a valve or the like.

Furthermore, a structure may be adopted in which the inside of the closed container is divided into a culture tank and a culture solution discharge tank using porous hollow fibers as a cell support.

Furthermore, as shown in FIG. 6, the cell support 1 may have a structure in which a porous membrane is folded into many layers.

Furthermore, the sealed container may be made of a transparent material.

Cell proliferation status and separation of cells from the support by backwashing
It may be possible to observe the floating state.

[Effects of the invention] As explained above, the cultured and proliferated cells can be separated from the cell support and suspended in a solution, making it possible to collect and recover the cells themselves. ■For the food and chemical fields where cells are used as bioreactors.■For the medical and pharmaceutical fields where cells, such as immunocompetent cells, are differentiated and proliferated outside the body and returned to the body to treat cancer, etc., stably and safely. Effective in supplying target cells.

[Brief explanation of the drawing]

FIG. 1 is a sectional view showing one embodiment of the cell culture container of the present invention, FIGS. 2 and 3 are sectional views showing the operation, and FIGS. It is a figure showing an example of. l... Cell support (porous membrane), 2... Sealed container,
3... Culture solution supply port, 4... Culture solution outlet, 5...
・Backwash liquid supply port, 6...Cell collection port, 7...Cells,
8...Culture solution, 9...Culture tank, 10...Culture solution discharge tank Patent applicant Sumitomo Electric Industries Co., Ltd. Agent Patent attorney Aohaku Ao (1 person) Figure 1 Figure 2 Figure 3 Figure 4 % formula % Name of the invention Cell culture container 3, relationship with the person making the amendment Case Patent applicant 1999? Month 130
Change of ward name/Change of residence indication by Ii & Address 4-5-33 Kitahama, Chuo-ku, Osaka-shi, Osaka Name (2)
13) Tetsu Kawakami, Representative of Sumitomo Electric Industries, Ltd.

Claims (2)

[Claims] (1) The inside of the airtight container is divided into a culture tank for culturing cells in a porous material and a culture solution discharge tank for discharging the culture solution that has passed through the porous material. A cell culture container characterized by a structure in which a collection port is connected to the culture solution discharge tank, and a culture solution discharge port and a backwash solution supply port are connected to the culture solution discharge tank. (2) The cell culture container according to claim 1, wherein a transparent material is used as the closed container.