JP2008136414A - Cell culture facility - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To effectively suppress a rise in humidity in a room in a cell culture facility using a liquid nitrogen tank. <P>SOLUTION: The cell culture facility 1 comprises a freezing preservation container 2 to which liquid nitrogen is supplied from a liquid nitrogen tank 31 through a liquid route 32 and a multi-gas incubator 6 for controlling the feed amount of a nitrogen gas and regulating a gas concentration, performs an operation related to a cell preparation, includes an air conditioner 21 for air-conditioning a room, an outdoor air treating unit 22 for treating outdoor air before introducing it into the air conditioner and a gas route 37 for supplying a nitrogen gas generated in the liquid nitrogen tank to the multi-gas incubator, and carries out heat exchange between the nitrogen gas passing through the gas route and the outdoor air treated by the outdoor air treating unit. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention is configured so that cells, microorganisms, bacteria, etc. (hereinafter collectively referred to as cells) are not leaked to the outside, and a cell culture in which a cryopreservation container for cryopreserving cells, an incubator for culturing cells, and the like are installed. It relates to a facility (CPC: Cell Processing Center).

  In recent years, research and development on regenerative medicine using cells has been promoted, and great expectations are placed on the results. In regenerative medicine, stem cells are proliferated by being compounded with various materials, and are guided to the form of a predetermined organ through various techniques such as differentiation induction and cell adhesion. Since the prepared organs and tissues are transplanted and transfused to individual patients, the cell culture operation is performed in an environment that satisfies the containment level defined in a predetermined standard, for example, GMP (Good Manufacturing Practice) standard. It is necessary to be done at

  Conventional cell culture facilities that perform cell culturing operations and the like have a room with a predetermined cleanliness, protect personnel and personnel engaged in work from viruses, and contain room air Thus, a facility that can prevent contamination of the surrounding environment has been proposed (see Patent Document 1).

  In this cell culture facility, as described above, cells that can be used for regenerative treatment are cultured, and aseptic control and aseptic operation are required. Therefore, in order to maintain the cleanliness of air, each room formed in the cell culture facility usually adopts the structure of a clean room that is used, and the air supply system for introducing outside air is used. Clean air is supplied by using a filter such as a medium performance filter or a high performance filter.

In addition, in this seed cell culture facility, in order to cryopreserve cells at an ultra-low temperature, a cryopreservation container for storing using a gas phase of liquid nitrogen is installed, and a multi-gas incubator and the like essential for cell culture are also provided. is set up. In this multi-gas incubator, the supply amount of nitrogen gas or carbon dioxide gas is controlled and adjusted to a predetermined gas concentration. Conventionally, a nitrogen gas cylinder has been used as a nitrogen gas source supplied to the multi-gas incubator.
JP 2003-47457 A JP 2006-81994 A

  Here, in the above-mentioned cell culture facility, in order to maintain the cleanliness of the air, the indoor air is circulated while being cleaned with a high-performance filter, the room temperature is adjusted with an air conditioner, and the outside air introduction amount is exhausted. Since it is used, the humidity in the room becomes high, and there is a concern that the working environment will deteriorate due to contamination. For this reason, an outside air processing unit that treats the outside air before being introduced into the indoor air conditioner is provided, and this outside air processing unit is designed to remove the moisture in the outside air as much as possible. In many cases, it was insufficient.

  On the other hand, in the liquid nitrogen tank for supplying liquid nitrogen to the cryopreservation container, nitrogen gas is constantly generated, so this liquid nitrogen tank is open to the atmosphere. Therefore, in Patent Document 2, the nitrogen gas discharged into the atmosphere is supplied to the multi-gas incubator so that the nitrogen gas can be effectively used, the nitrogen gas cylinder can be downsized, and the amount of gas used can be suppressed. Yes.

  The present invention has been made in view of the above-described conventional situation, and an object thereof is to effectively suppress an increase in indoor humidity in a cell culture facility that uses a liquid nitrogen tank.

  The cell culture facility of the invention of claim 1 is provided with a cryopreservation container in which liquid nitrogen is supplied from a liquid nitrogen tank through a liquid path, and an incubator in which the gas concentration is adjusted by controlling the supply amount of at least nitrogen gas This is a facility where work related to cell adjustment is carried out, including an air conditioner that air-conditions the room, an outside air treatment unit that treats the outside air before it is introduced into the air conditioner, and nitrogen gas generated in the liquid nitrogen tank. And a nitrogen gas flowing through the gas path and the outside air to be processed by the outside air processing unit are heat-exchanged.

  The cell culture facility of the invention of claim 2 is a facility in which a cryopreservation container to which liquid nitrogen is supplied from a liquid nitrogen tank via a liquid supply path is installed, and work related to cell adjustment is performed, and the room is air-conditioned. An air conditioner, an outside air processing unit for treating the outside air before introducing it into the air conditioner, and a gas path for discharging the nitrogen gas generated in the liquid nitrogen tank, and nitrogen gas flowing through the gas path, It is characterized by exchanging heat with the outside air processed by the outside air processing unit.

  According to the invention of claim 1, a cryopreservation container in which liquid nitrogen is supplied from a liquid nitrogen tank via a liquid path, and an incubator in which the gas concentration is adjusted by controlling the supply amount of at least nitrogen gas are installed, In cell culture facilities where work related to cell conditioning is performed, an air conditioner that air-conditions the room, an outside air treatment unit that treats the outside air before it is introduced into the air conditioner, and nitrogen gas generated in the liquid nitrogen tank is supplied to the incubator A gas path for supplying the nitrogen gas in the liquid nitrogen tank that has been simply discharged to the incubator for effective use, and reducing the size of the nitrogen gas cylinder that is normally installed in the incubator, Nitrogen consumption can be reduced.

  In particular, since the nitrogen gas flowing through the gas path for supplying the nitrogen gas to the incubator and the outside air to be processed by the outside air processing unit are heat-exchanged, the humidity in the outside air introduced into the room is changed to the gas path. It can be cooled and condensed by the cryogenic nitrogen gas flowing inside and removed. As a result, an increase in indoor humidity of the cell culture facility can be effectively suppressed, and a clean and comfortable working environment can be realized.

  In addition, since the temperature of the nitrogen gas supplied to the incubator can be increased, the adverse effect on the culture environment caused by the supply of low-temperature nitrogen gas to the incubator can be eliminated.

  According to the second aspect of the present invention, in a cell culture facility where a cryopreservation container to which liquid nitrogen is supplied from a liquid nitrogen tank via a liquid supply path is installed and work related to cell adjustment is performed, air conditioning for air conditioning the room And an outside air treatment unit for treating the outside air before introducing it into the air conditioner, and a gas path for discharging the nitrogen gas generated in the liquid nitrogen tank. Heat exchange with the outside air processed in the unit allows the moisture in the outside air introduced into the room to be cooled and condensed by the cryogenic nitrogen gas flowing in the gas path and removed. It becomes like this. As a result, it is possible to effectively use the nitrogen gas in the liquid nitrogen tank that has been simply exhausted so as to effectively suppress an increase in the room humidity of the cell culture facility and realize a clean and comfortable working environment. become.

  Hereinafter, an embodiment of the present invention will be described in detail with reference to the drawings. FIG. 1 is a schematic explanatory view for explaining the liquid nitrogen supply, nitrogen gas supply, and air conditioning system of the cell culture facility 1 to which the present invention is applied, and FIG. 2 is a plan view for explaining the layout of the cell culture facility 1 as a whole. Show.

  In FIG. 2, the cell culture facility 1 is installed in a building that is a site of regenerative medicine or in vitro fertilization medicine, such as a university laboratory, a hospital, and a target specimen in regenerative medicine or in vitro fertilization medicine. A plurality of cell cultures in which a cryopreservation container 2 for storing cryopreserved cells (for example, donor cells), a cell storage chamber 4 in which an ultra-low temperature freezer 3 is installed, and a multi-gas incubator 6 for performing cell culture are installed In addition to the chamber 7..., An entrance chamber 8, a monitoring chamber 9, a primary chambering chamber 11, a supply chamber 12, an isolator chamber 13, a secondary chambering chamber 14, a degauning chamber 16 and the like are defined.

  Each room is distinguished by four grades of air cleanliness, A to D. Grade D has the highest cleanliness less than CLASS100, Grade C has the second highest cleanliness after D and less than CLASS10000, Grade B The cleanliness is next to C and is less than CLASS100,000, and Grade A has the lowest cleanliness and is CLASS100000 or more (see FIG. 2). The air cleaning and air conditioning in each room is performed by the air conditioner 21 for each room and the outdoor air processing unit 22 in the previous stage as described later. In addition, the layout of each room is designed to minimize changes in contamination and air cleanliness in consideration of the flow of people and things.

  The multi-gas incubator 6 installed in the cell culture chamber 7 is configured with a heat insulation box having an opening on the front surface and a heat insulation door for closing the opening so as to be openable and closable. A cell (sample) to be cultured is cultured while maintaining a constant gas concentration such as temperature, carbon dioxide concentration, oxygen concentration, etc., and the inside is aseptic. Nitrogen gas is used to control this gas concentration. In addition, when culturing cells, a heater and a control device for adjusting the temperature in the culture chamber are provided. The inside of this culture chamber is divided vertically by a plurality of shelves, and a humidifying dish in which water for humidification is stored is disposed on the bottom surface of the culture chamber, and is provided by a heater provided outside the stainless steel inner box. Heat to evaporate water. An inner door is provided inside the heat insulating door.

  In the cell culture chamber 7, in addition to the multi-gas incubator 6, a medical cool box 23 that keeps the medicine cool, a biohazard cabinet 24 in which the working space is less than CLASS 100, a centrifuge 26, and the like are installed.

  Outside the cell culture facility 1, a liquid nitrogen tank 31 for storing liquid nitrogen having a boiling point of -195.8 ° C. by applying pressure is installed, and a liquid path (from a vacuum insulation pipe) is inserted to the bottom of the tank. And the cryopreservation container 2 and the ultra-low temperature freezer 3 in the cell storage chamber 4. Since the liquid nitrogen is always spontaneously evaporated in the liquid nitrogen tank 31, the internal pressure is high, and the liquid nitrogen is pushed out to the liquid path 32 by this evaporation pressure, and each cryopreservation container 2 and the ultra-low temperature freezer 3. The liquid nitrogen is conveyed to the liquid nitrogen tank (the liquid nitrogen tank 31 is provided with a pressure valve (safety valve) (not shown) that opens to the atmosphere above a predetermined dangerous pressure). In this case, a valve 33 is provided in one of the branched liquid paths 32, and the automatic supply control device 36 controls the valve 33 so that the liquid nitrogen from the liquid nitrogen tank 31 to each cryopreservation container 2 is controlled. The supply of (LN2) is controlled. That is, the automatic supply control device 36 controls the valve 33 to control the liquid nitrogen level in the cryopreservation container 2 to a predetermined value. The cryopreservation container 2 is composed of a vacuum heat insulating container, and the cells (samples) housed inside are cryopreserved at about −190 ° C. by evaporated cryogenic nitrogen gas.

  Further, a valve 34 is interposed in the other branched liquid path 32, and the supply of liquid nitrogen from the liquid nitrogen tank 31 to the ultralow temperature freezer 3 is controlled by the ultralow temperature freezer 3 controlling the valve 34. Is done. This ultra-low temperature freezer 3 cools the storage chamber to an extremely low temperature of −150 ° C. or lower by a multi-source multi-stage refrigerant circuit equipped with a compressor. When a failure occurs in this refrigerant circuit, the ultra-low temperature freezer 3 opens the valve 34. Supply liquid nitrogen into the storage chamber. Thereby, it is comprised so that the emergency can be coped with by maintaining the storage room at a very low temperature.

  Although the ultra-low temperature freezer 3 has conventionally been provided with a separate liquid nitrogen tank for dealing with such abnormal situations, liquid nitrogen can be supplied from the liquid nitrogen tank 31 for the cryopreservation container 2 as in the embodiment. As a result, it is no longer necessary to use a liquid nitrogen tank for the ultra-low temperature freezer.

  Here, when the outdoor temperature is considered to be an average of + 20 ° C., the temperature difference between the inside and outside of the liquid nitrogen tank 31 is actually about 220 deg. Therefore, the liquid nitrogen in the liquid nitrogen tank 31 is always naturally evaporated as described above. In addition, if the pressure in the liquid nitrogen tank 31 increases too much, there is a danger of explosion, so when the pressure exceeds the predetermined pressure, it is necessary to periodically discharge the nitrogen gas generated by this natural evaporation. .

  Therefore, in the present invention, a gas path 37 that communicates between the upper part of the liquid nitrogen tank 31 and each multi-gas incubator 6 is provided. The gas path 37 further communicates with the upper part of the cryopreservation container 2 so that nitrogen gas generated by evaporation inside both the liquid nitrogen tank 31 and the cryopreservation container 2 flows. And the gas concentration in each multi-gas incubator 6 is controlled by controlling the nitrogen gas supply amount from this gas path 37 by the control apparatus of the multi-gas incubator 6 mentioned above.

  As a result, the nitrogen gas in the liquid nitrogen tank 31 or the nitrogen gas in the cryopreservation container 2 that has been regularly discharged until now is supplied to the multi-gas incubator 6 through the gas path 37 to effectively use the nitrogen gas. Therefore, it is possible to reduce the size and the amount of nitrogen used in a nitrogen gas cylinder filled with a compressed nitrogen gas that is always provided in the multi-gas incubator 6.

  A pressure valve 38 is provided in the middle of the gas path 37 so that when the pressure of the nitrogen gas rises above a predetermined value, the gas path 37 is discharged from the branch path 37A of the gas path 37 to the outside. In addition, a backup nitrogen gas cylinder 40 is connected to the downstream side of the heat exchanger 37B (described later) of the gas path 37.

  Next, the air conditioning system of the cell culture facility 1 will be described. The outside air processing unit 22 is provided with a pre-filter 41 that removes relatively large dust, a blower 42, a medium-performance filter 43 that removes relatively small dust, and a heat exchanger 44. The heat exchanger 44 constitutes an evaporator of a refrigerant circuit (not shown) provided with a compressor. Further, a heat exchanger 37B constituting a part of the gas path 37 is arranged on the air suction side of the outside air processing unit 22 so as to exchange heat with outside air sucked into the outside air processing unit 22 by the blower 42. It is configured. The gas path 37 leading to the heat exchanger 37B is thermally insulated by a heat insulating material 37C. Reference numeral 46 denotes a reheat heater (electric heater) that heats the air emitted from the outside air processing unit 22.

  The air (outside air) that has passed through the outside air processing unit 22 and the reheat heater 46 enters the air conditioners (showing the air conditioner of the cell culture chamber 7 in FIG. 1) 21 through the duct 47. The air conditioner 21 includes a blower 48, a heat exchanger 49, and a HEPA filter (high performance filter) 51. The heat exchanger 49 constitutes an evaporator or condenser of a refrigerant circuit (not shown) provided with a compressor. The blower 48 sucks air in the cell culture chamber 7 from the duct 52 together with air from the duct 47 (outside air), and discharges it into the cell culture chamber 7 through the heat exchanger 49 and the HEPA filter 51. In addition, the air in the cell culture chamber 7 is further discharged to the outside by an exhaust fan 54 via a duct 53.

  With the above configuration, when the blower 42 is operated, outside air is introduced into the outside air processing unit 22. The introduced outside air exchanges heat with a heat exchanger 37B provided in the gas path 37. Since the cryogenic nitrogen gas circulates in the heat exchanger 37B as described above, moisture in the outside air exchanged with the heat exchanger 37B condenses and adheres to the surface of the heat exchanger 37B as condensation. . Since this dew condensation is discharged through a drainage passage (not shown), the outside air introduced into the outside air processing unit 22 is dehumidified, and its absolute humidity is significantly reduced.

  The dehumidified outside air is further condensed and removed by being cooled by the heat exchanger 44 after dust is removed by the pre-filter 41 and the medium performance filter 43. Then, it is sent to the reheat heater 46, heated to a temperature of about + 22 ° C., and then sent to the air conditioner 21. When the blower 48 is operated, air (outside air) from the reheat heater 46 is introduced into the heat exchanger 49, and after removing microscopic dust and viruses with the HEPA filter 51, the cell culture chamber 7 It is discharged inside. The air in the cell culture chamber 7 is further sucked into the blower 48 through the duct 52 and is repeatedly returned to the cell culture chamber 7 through the heat exchanger 49 and the HEPA filter 51. Thereby, the inside of the cell culture chamber 7 is maintained at the above-described cleanliness, and the temperature is maintained at a predetermined temperature by heat exchange with the heat exchanger 49.

  The blower 54 discharges the air in the cell culture chamber 7 at a constant air speed. By adjusting this exhaust and the amount of outside air introduced from the outside air processing unit 22, in the embodiment, the left and right cell culture chambers 7 in FIG. For example, the chamber pressure is controlled to +30 Pa, and the central cell culture chamber 7 is controlled to −10 Pa. The room force is not limited to this, and may be controlled to 0 Pa (atmospheric pressure), for example, depending on the purpose of use. Although only the cell culture chamber 7 is shown in FIG. 1, the air conditioning in the other chambers is the same. In the examples, the cell storage chamber 4 is −10 Pa, the entrance chamber 8 is 0 Pa, the monitoring chamber 9 is also 0 Pa, the primary chambering chamber 11 is +5 Pa, the supply chamber 12 is +15 Pa, the isolator chamber 13 is +20 Pa, the secondary chambering chamber 14 is +15 Pa, The degauning chamber 16 is set to +10 Pa.

  Here, the cell culture chamber 7 and the like maintain its cleanness while circulating air as described above, and since the outside air is introduced in an amount substantially equal to the displacement, the humidity is inevitably increased, such as a scallop insect and a fungus. The problem of breeding occurs. However, in the present invention, as described above, the heat exchanger 37B in the middle of the gas pipe 37 through which the nitrogen gas generated in the liquid nitrogen tank 31 and the cryopreservation container 2 flows is arranged on the air introduction side of the outside air processing unit 22, and a multi-gas incubator. Since the nitrogen gas supplied to 6 and the outside air processed by the outside air processing unit 22 are heat-exchanged, the moisture in the outside air introduced into the cell culture chamber 7 is used as the pole that flows in the gas path 37. It becomes possible to cool and condense with a low-temperature nitrogen gas and remove it. As a result, it is possible to effectively suppress an increase in humidity in the cell culture room 7 of the cell culture facility and other rooms, and to realize a clean and comfortable working environment.

  In particular, the temperature of the nitrogen gas supplied to the multi-gas incubator 6 rises by absorbing heat from the outside air in the heat exchanger 37B, so that low-temperature nitrogen gas is supplied to the multi-gas incubator 6 and the internal temperature decreases. It is also possible to prevent inconvenience.

  In the embodiment, the multi-gas incubator 6 is used as an incubator. However, the present invention is not limited to this, and any incubator that controls the gas concentration using nitrogen gas may be used. In the embodiment, the case where nitrogen gas is supplied from the liquid nitrogen tank 31 or the like to the multi-gas incubator 6 has been described. However, the invention of claim 2 is not limited thereto, and the nitrogen gas is simply discharged from the branch path 37A (multi-gas). Even without the supply to the incubator 6), it is possible to effectively remove the moisture in the outside air.

It is a schematic explanatory drawing explaining the liquid nitrogen supply of the cell culture facility to which this invention is applied, nitrogen gas supply, and an air-conditioning system. It is a top view explaining the layout of the whole cell culture facility of FIG.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Cell culture facility 2 Cryopreservation container 3 Ultra-low temperature freezer 4 Cell storage room 6 Multi gas incubator 7 Cell culture room 21 Air conditioner 22 Outside air processing unit 31 Liquid nitrogen tank 32 Liquid path 37 Gas path 37B Heat exchanger

Claims (2)

A cell in which work related to cell adjustment is performed by installing a cryopreservation container in which liquid nitrogen is supplied from a liquid nitrogen tank through a liquid path and an incubator in which at least the supply amount of nitrogen gas is controlled to adjust the gas concentration A culture facility,
An air conditioner that air-conditions the room;
An outside air processing unit for treating outside air before introducing it into the air conditioner;
A gas path for supplying nitrogen gas generated in the liquid nitrogen tank to the incubator,
A cell culture facility characterized in that heat exchange is performed between nitrogen gas flowing through the gas path and outside air to be treated by the outside air treatment unit. A cell culture facility in which a cryopreservation container to which liquid nitrogen is supplied from a liquid nitrogen tank via a liquid supply path is installed, and work related to cell adjustment is performed,
An air conditioner that air-conditions the room;
An outside air processing unit for treating outside air before introducing it into the air conditioner;
A gas path for discharging the nitrogen gas generated in the liquid nitrogen tank,
A cell culture facility characterized in that heat exchange is performed between nitrogen gas flowing through the gas path and outside air to be treated by the outside air treatment unit.

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