JP3095464B2 - Cell culture device - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus for culturing cells such as animal cells in a liquid.

[0002]

2. Description of the Related Art Conventionally, there has been an apparatus for continuously and long-term culturing cells, but a membrane for separating cells from a circulating medium has not been used.

[0003]

In the prior art, since a separation membrane is not built in the cell culture vessel, the cells spread over the entire medium circulation pipe, and oxygen supply and carbon dioxide removal are not performed efficiently . Furthermore, a separation membrane is placed inside the cell culture vessel.
Built-in to prevent cells from spreading throughout the media circulation piping
To efficiently supply oxygen and remove carbon dioxide gas,
Because fresh medium flows directly into the cell culture room
Depending on the cells to be cultured in
There is a problem that the generated fluid force causes serious damage to the cells .

According to the present invention, a cell culture vessel is divided into two chambers by a separation membrane, and one of the chambers is generated by a discharge pressure of a pump.
Environment that is not suitable for cell culture
The cell culture chamber that can be stationary state that the medium and the other chamber
As medium circulation chamber for passing, so as not cells spread into the medium circulation chamber, as well as efficient oxygen supply and carbon dioxide removal to the medium carried out in circulation process, cells in culture
It is an object of the present invention to provide a cell culture device capable of further reducing damage to cells.

[0005]

SUMMARY OF THE INVENTION The present invention is to provide a cell culture apparatus described in the following (1) and (2) as means for solving the above-mentioned problems. (1) A medium outlet and inlet are provided, and the medium is circulated.
A medium circulation chamber and a medium circulation chamber,
Circulating medium of the water substance and the cell culture vessel which is partitioned into two chambers with the transmitted cells cell culture chamber separation membrane impermeable is stretched, the medium before Symbol medium circulation chamber between the medium circulation chambers Let
A medium circulation piping which pump is interposed that the oxygenator and carbon dioxide remover for performing each carbon dioxide removal from the oxygen supply Kyu及 beauty culture into the medium interposed in the medium circulation pipe,
Before SL communicates with the medium circulation piping, cell culture apparatus characterized by comprising; and a medium container for replenishing the culture <br/> locations on the medium circulating in the pipe. (2) in cell culture apparatus according to (1), before
For sampling or sample injection or the like in the serial cell culture chamber,
A cell culture apparatus comprising a port penetrating the cell culture chamber .

[0006]

Since the present invention is constructed as described above, it has the following functions (1) and (2). (1), in the configuration of the above (1) <br/> water substances to the cell culture vessel is transmitted, the separation membrane cell does not transmit, partition into two chambers of the cell culture chamber and the medium circulating chamber Since the medium circulation pipe is disposed in the medium circulation chamber, cells do not enter the medium circulation chamber, and therefore, cells do not flow through the circulation pipe, so that oxygen supply and carbon dioxide removal performed in the medium circulation pipe are efficient. It is done on a regular basis. In addition, medium circulation in the cell culture room
Water is simply diffused from the annulus through the separation membrane.
Since the ground does not flow directly into the cell culture room,
Culture in a cell culture room without disturbing the culture
Cells are not damaged by these disturbances.
No. In addition to the pump, an oxygen supply and a carbon dioxide gas remover are interposed in the medium circulation pipe, and water-soluble nutrients and cell wastes, which are the medium in the cell culture room, are separated into the medium circulation room. Since the liquid can freely enter and exit through the membrane, the liquid flowing through the circulation pipe becomes oxygen-rich and decarbonated, that is, returns to the medium circulation chamber in a state favorable for cell culture,
As a result, the medium is enriched in oxygen-rich and dilute waste products as it passes through the separation membrane and enters the cell culture chamber. In addition, since the medium container is connected to the medium circulation pipe, the medium can be automatically replenished as needed. (2) In the above configuration (2), in addition to the configuration of the above (1), the cell culture chamber is provided with a port for sample collection / injection, etc., so that cell culture can be performed by collecting a sample when necessary. Observation of the state of cell culture in the room, injection of nutrient solution and the like can be easily performed.

[0007]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A first embodiment of the present invention will be described with reference to FIG.
FIG. 1 is a schematic configuration diagram of the present embodiment. In the figure, a cell culture vessel 1 is divided into a cell culture chamber 3 and a medium circulation chamber 4 by a separation membrane 2. The medium circulation chamber 4 is provided with a pump 8 and a medium circulation pipe 5 having a required valve (not shown). The medium circulation pipe 5 is provided with an oxygen supplier 6 and a carbon dioxide gas remover 7, and after exiting the medium circulation chamber 4, the medium maintained at the appropriate dissolved oxygen concentration and dissolved carbon dioxide concentration by these means The medium is again supplied to the culture medium circulation chamber 4 by the pump 8, and the same amount of culture medium is discharged from the culture medium circulation chamber 4 and sent to the oxygen supply device 6 and the carbon dioxide gas removal device 7. In this way, the dissolved oxygen and dissolved carbon dioxide in the medium circulation chamber 4 are maintained at appropriate concentrations. A medium container 9 is connected to the medium circulation pipe 5 through a valve (not shown). The valve is opened and closed as necessary, and fresh medium is supplied to the medium circulation chamber 4 through the medium circulation pipe 5 and is cultured. The same amount of medium containing accumulated waste is discharged to the waste liquid container 10. In this way,
Nutrient components and waste components in the medium circulation chamber 4 are always kept at a constant level. The separation membrane 2 that divides the cell culture chamber 3 and the medium circulation chamber 4 does not allow the cells 11 to pass through but allows the medium components to pass through (eg, a membrane filter). Therefore, the medium component in the cell culture room 3 and the medium component in the medium circulation room 4 move mutually via the separation membrane 2, and the medium in the cell culture room 3 is almost the same as the fresh medium in the medium circulation room 4. Condition can be maintained. As described above, according to the cell culture apparatus of the present embodiment, the cell medium is circulated through the medium circulation pipe 5 and the medium is always kept fresh by passing through the oxygen supply device 6 and the carbon dioxide gas remover 7, so that the cells 11 can be kept for a long time. Continuous culture can be performed. In cell culture, cells are not simply cultured for a long time in a free state, but involve the collection of metabolites and observation of the cell culture state. In addition, special reagents may be injected during culture. Thus, FIG. 2 shows a schematic configuration diagram of the cell culture device of the second embodiment configured to be suitable for these controls. That is, by attaching the sampling port 12 to the cell culture room 3, it is possible to adapt to these management operations. Other configurations are the same as in the first embodiment.

The cell culture vessel 1 can be made transparent or not, but it is possible to observe the cell culture state more clearly by making it transparent or by providing a transparent window. Therefore, a more appropriate culture state can be maintained.

As described above, according to the first and second embodiments,
A water permeable but cell-impermeable separating membrane 2 is used to partition the inside of the cell culture vessel 1. One of the chambers is used as the cell culture chamber 3 and the other is used as the medium circulation chamber 4.
Is provided with a medium circulation pipe 5 provided with an oxygen supply device 6 and a carbon dioxide gas removal device 7, and circulated by a pump 8, so that cells in the cell culture chamber 3 do not enter the circulating fluid.
There is an advantage that the circulating fluid, that is, the medium is efficiently enriched with oxygen and decarbonated, and as a result, cells can be stably cultured for a long period of time.

[0010]

Since the present invention is configured as described above,
Cell culture media can always be kept fresh, and
The cells can be cultured in a stable state for a long time without damaging the cells.

[Brief description of the drawings]

FIG. 1 is a schematic configuration diagram of a cell culture device according to a first embodiment of the present invention.

FIG. 2 is a schematic configuration diagram of a cell culture device according to a second embodiment of the present invention.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Cell culture container 2 Separation membrane 3 Cell culture room 4 Medium circulation room 5 Medium circulation pipe 6 Oxygen supply device 7 Carbon dioxide gas remover 8 Pump 9 Medium container 10 Waste liquid container 11 Cell 12 Sampling port

──────────────────────────────────────────────────続 き Continuing from the front page (72) Inventor Shinichi Shinozaki 1-1-1, Wadazakicho, Hyogo-ku, Kobe-shi Inside Kobe Shipyard, Mitsubishi Heavy Industries, Ltd. (72) Inventor Kazuhide Uemura 5-chome Komatsudori, Hyogo-ku, Kobe-shi No. 1-16 Inside Shinryo Hitec Co., Ltd. (56) References JP-A-2-13365 (JP, A) WO 90/190 (WO, A1) (58) Fields investigated (Int. Cl. ⁷ , (DB name) C12M 1/00-3/06

Claims (2)

(57) [Claims] 1. An outlet and an inlet for a culture medium are provided to circulate the culture medium.
And a medium circulation chamber to be disposed and adjacent to the medium circulation chamber.
Is a cell culture vessel which medium water material permeable separating membrane cells is not transmitted is divided into two chambers with cell culture chamber stretched between the medium circulation chamber, before Symbol medium circulation chambers oxygen performing the medium circulation piping which pump causes <br/> circulating medium is interposed, carbon dioxide removal from the oxygen supply Kyu及 beauty culture into the medium interposed in the culture <br/> locations circulation piping, respectively a feeder and carbon dioxide remover, communicated before Symbol medium circulation piping, wherein the medium circulation distribution
A cell culture device, comprising: a medium container for replenishing a medium in a tube . In cell culture apparatus according to claim 1, such as for sampling or sample injection in the cell culture chamber, cells, characterized by comprising comprises a port that penetrates into the cell culture chamber Culture device.