JP2007074921A - Cell culture device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cell culture device enabling safe culture while preventing the intrusion (contamination) of foreign materials during the cell culture. <P>SOLUTION: The cell culture device is provided with a plurality of culture vessels 2, 4 having a charging port and a discharging port and made of a gas-permeable material, and a gastight casing 20 holding the multiple culture vessels. The culture vessel and the casing 20 are formed in cylindrical form and the casing 20 is rotated around the center axis with a rotating means 90. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a cell culture apparatus, and more particularly, to a culture dish structure that can reduce the risk of contamination and can be densified.

In cell culture, complicated passage processes such as replacement of the medium in the incubator and re-seeding to make the cell density appropriate are performed manually. Usually, these operations are carefully performed in a relatively clean atmosphere in which airborne particulates are reduced by a clean environment generation technique cultivated in the semiconductor manufacturing field in order to avoid the occurrence of contamination. However, even in this clean atmosphere, it is not sufficient to completely avoid contamination. Usually, a circular petri dish used as an incubator (observing cells appropriately with a microscope to measure the passage timing described later during culture) However, when culturing cells using a petri dish, particularly in adherent cells), be careful not to mix bacteria by lifting the petri dish up so that the lid of the petri dish is lifted when changing the medium. It was necessary to perform complicated and difficult work such as working quickly while paying attention to the pipettor in the gap between the petri dish and its lid and not touching the surroundings such as the edge of the petri dish. (For example, Patent Document 1)
JP 2004-16194 A

  As described above, the culturing work is currently performed manually despite being cumbersome, and it is difficult to perform easily because it requires skilled work. .

  In the following, further specific examples will be used to explain the development of technology for regenerative medicine. In the case of stem cell culture for constructing tissues, the cultured cells are transplanted into a subject. Therefore, the possibility of contamination (contamination) during culture must be guaranteed at 0%. However, the above-mentioned clean environment generation technology reduces the concentration of suspended particles in the atmosphere, but various electrical and mechanical parts are usually arranged in the environment, and substantially fine particles. It is extremely difficult to guarantee the number of 0. The requirement for avoiding contamination required for cell culture is that the number of living bacteria mixed in the medium should be zero, and even if it is one, contamination with live bacteria It becomes. As described above, the clean environment generation technology is effective in reducing the risk, but there is a big problem that it is not possible to guarantee avoidance of contamination. In addition, culturing a plurality of subject cells close to each other, specifically in one room, increases the risk in terms of cross-contamination. When fungi (for example, mold) are mixed in a subject's cells for some reason, the possibility that the spores will spread and be transmitted to other subject cells increases. In addition, when trying to obtain a larger number of cells by culturing for a longer period of time, it is often accompanied by an operation called passage. In this operation, since the growth is inhibited when the density of the cultured cells is increased, the cells are newly separated and seeded again in a plurality of petri dishes, and the number of petri dishes is increased. As the number of petri dishes increases, a larger temperature chamber is required for culturing, which inevitably increases the risk of contamination. As described above, the conventional cell culture work has a big problem in safety.

  The cell culture apparatus of the present invention includes a plurality of incubators having an inlet and an outlet and formed of a gas permeable material, and an airtight casing including the plurality of incubators. Further, the incubator and the casing are formed in a cylindrical shape, and are provided with rotating means for rotating the casing around the central axis.

  ADVANTAGE OF THE INVENTION According to this invention, the cell culture apparatus which can avoid the risk of contamination can be provided.

  Hereinafter, an embodiment of a cell culture device to which the present invention is applied will be described with reference to FIG. 1 and FIG. FIG. 1 is a block diagram showing a schematic configuration of a cell culture apparatus to which the present invention is applied. Container 2 is a primary incubator, containers 4 and 16 are secondary incubators, and containers 6, 8, 10, 12, 14, and 18 are third incubators. The envelope of each of these incubators is formed of a known gas permeable material. The case 20 includes the primary incubator, the second incubator, and the third incubator. External air flows into the housing 20 through the pump 54 and the filter 52, and the air inside the housing 20 is discharged to the outside through the filter 56. Note that these filters 52 and 56 prevent large dust and dust from entering the inside of the housing 20 and prevent an image from being captured by an image acquisition unit to be described later. The containers 40, 42, and 44 are infusion bags for storing chemicals necessary for cell culture. When the cultured cells are adhesion-dependent, they are usually a medium, a cell release agent, and a buffer, respectively. The container 58 is a waste solution bag for storing a buffer solution and a cell detachment agent used in the culture process, and a used medium which is usually changed every two or three days. The above infusion bags 40, 42, 44, primary incubator 2, second incubator 4, 16, 3rd incubator 6, 8, 10, 12, 14, 18, and waste solution bag 58 are connected by a tube (shown by a solid line). Do not remove the tube during culture. The pinch valves 34, 36, 38, 22, 24, 26, 46, 48, and 50 control liquid feeding. The pumps 28, 30, 32, and 60 are preferably peristaltic pumps (known pumps such as squeezing pumps and roller pumps) in order to prevent the liquid from touching the outside.

  FIG. 2 is a detailed view of the incubator of the cell culturing apparatus to which the present invention is applied, and FIG. 2 (a) shows the relationship between the incubator and the units provided around it. A cross-sectional view and FIG. 2 (b) are perspective views of the incubator. A cylindrical incubator 2, 4, 6, 8, 10, 12, 14, 16 is contained in the cylindrical housing 20. The circumferential sides of these incubators 2, 4, 6, 8, 10, 12, 14, and 16 are made of gas permeable material, and the inlet and outlet are formed at both ends, and tubes are connected to them. ing. The connection of these tubes is shown in FIG. A light source 70 and image acquisition means 72 including a lens and an image sensor are disposed above and below the housing 20. The image acquisition means 72 is held by a vertical movement means 82. The vertical movement means 82 is connected to a lateral movement means comprising a motor 80 and pulleys 74, 78 and a belt 76, and can be slid in the C direction by driving the motor 80. . The vertical movement means 82 is similarly configured by a known technique including a pulley, a belt, and a motor. The light source 70 and the motor 0 are connected to the controller 84. A motor 90 that drives the rollers 86 and 88 is connected to the controller 84. As a result, the casing 20 can rotate around the central axis L (rotation operation means).

1. Next, the culture process of the Example of this invention is demonstrated.
Step 1: {Cell injection} The pinch valve 22 is opened, and the pump 28 is driven to inject a liquid containing cells with a syringe from a branch portion of the tube. (Cell in)

  Step 2: {Medium injection} The pinch valves 34 and 22 are opened, and the pump 28 is driven to inject the culture medium into the incubator 2.

Step 3: {Medium discharge} The pinch valves 46 and 51 are opened, and the pump 60 is driven to discharge the culture medium.
* Repeat steps 2 and 3 as appropriate (eg every 3 days).

  Step 4: {Passage} The pinch valves 22 and 38 are opened, and the pump 28 is driven to inject the buffer into the incubator 2. The pinch valves 46 and 51 are opened, and the pump 60 is driven to discharge the buffer solution. The pinch valves 22 and 36 are opened, and the pump 28 is driven to inject the cell detachment agent into the incubator 2. The pinch valves 22 and 24 are opened, and the pumps 28 and 30 are driven to transfer the solution in the incubator 2 to the incubators 4 and 16.

  Step 5: {Medium injection} The pinch valves 34 and 24 are opened, and the pump 30 is driven to inject the culture media into the incubators 4 and 16.

Step 6: {Medium discharge} The pinch valves 48 and 51 are opened, and the pump 60 is driven to discharge the culture medium.
Repeat steps 5 and 6 as appropriate (eg every 3 days).

  Step 7: {Passage} The pinch valves 24 and 38 are opened, and the pump 30 is driven to inject the buffer into the incubators 4 and 16. The pinch valves 48 and 51 are opened, and the pump 60 is driven to discharge the buffer solution. The pinch valves 24 and 36 are opened, and the pump 30 is driven to inject the cell detachment agent into the incubators 4 and 16. The pinch valves 24 and 26 are opened, and the pumps 30 and 32 are driven to transfer the solution in the incubators 4 and 16 to the incubators 6, 8, 10, 12, and 14.

  Step 8: {Medium injection} The pinch valves 26 and 34 are opened, and the pump 32 is driven to inject the culture media into the incubators 6, 8, 10, 12, and 14.

Step 9: {Medium discharge} The pinch valves 50 and 51 are opened, and the pump 60 is driven to discharge the culture medium.
Repeat steps 8 and 9 as appropriate (eg every 3 days).

Step 10: {Recovery 1} The pinch valves 26 and 34 are opened, and the pump 32 is driven to discharge the cells.
Step 10: {Recovery 2} The pinch valve 50 is opened (the pinch valve 51 remains closed), and the pump 60 is driven to discharge cells (to Cell out).

During the culture, observation by the image acquisition means 72 is appropriately performed. The observation position can be moved in the direction indicated by the arrow C by the slide means described above. Further, the incubator to be observed can be changed by the above-described vertical movement means and the rotation operation means.
In the above description, the number of incubators can be changed. In addition, the number of passages can be changed.

  ADVANTAGE OF THE INVENTION According to this invention, the cell culture apparatus which can avoid contamination in cell culture is realizable. In addition, even when many cells are cultured, the incubator can contribute to downsizing that enables image observation.

The block diagram which shows schematic structure of the cell culture apparatus to which this invention is applied. The detailed drawing of the mechanism part of the cell culture device formed by applying the present invention.

Explanation of symbols

  2,4.6,8,10,12,14,16 Incubator, 20 housing, 72 Image acquisition means

Claims (2)

  A cell culture apparatus comprising: a plurality of incubators having an inlet and an outlet and formed of a gas-permeable material; and an airtight housing including the plurality of incubators.   A cell culture apparatus, wherein the incubator and the casing are formed in a cylindrical shape, and the rotating means rotates the casing about a central axis.

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