JP2010142196A - Cell culture apparatus and cell culture cassette - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cell culture apparatus which can reduce the amount of a reagent left in supply piping, and to provide a cell culture cassette. <P>SOLUTION: This cell culture apparatus 100 comprises reagent containers 210 for receiving reagents at a cold reserving temperature T<SB>1</SB>, a cell culture container 231 for receiving culture cells at a higher constant temperature T<SB>2</SB>than the cold reserving temperature T<SB>1</SB>, supply piping 240 for constituting a passage for the reagents supplied from the reagent containers 210 to the cell culture container 231, and a heating portion 21 for heating the surface of the supply piping 240 to a prescribed temperature T<SB>3</SB>between the reagent containers 210 and the cell culture container 231. The prescribed temperature T<SB>3</SB>is higher than the constant temperature T<SB>2</SB>. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a cell culture apparatus and a cell culture cassette for culturing cells.

  Conventionally, a cell culture device provided with a culture dish, a flask, or the like is known as a cell culture container that contains a medium for culturing cells (for example, Patent Document 1).

  Specifically, the cell culture apparatus includes a first reagent container that stores a small amount of reagent and a second reagent container that stores a large amount of reagent. The cell culture apparatus connects the first reagent container and the cell culture container, and has a first supply pipe for supplying a small amount of reagent from the first reagent container to the cell culture container. Similarly, the cell culture device connects the second reagent container and the cell culture container, and has a second supply pipe for supplying a large amount of reagent from the second reagent container to the cell culture container.

By the way, the insides of the first reagent container and the second reagent container are kept at a cold temperature T ₁ (for example, 4 ° C.). On the other hand, the inside of the cell culture container is kept at a constant temperature T ₂ (37 ° C.). Therefore, in order to reduce the damage of cells in cell culture vessel is already cultivated, the reagent from the first reagent container and the second reagent container when supplying to the cell culture vessel, the temperature of the reagent thermostatic temperature T ₂ It needs to be close.

Here, for the small amounts of reagents, by lengthening the first supply pipe, close to the temperature of the small amounts of reagents in thermostatic temperature T _2. For a large amount of reagent, a heat insulation for raising the temperature of the large amount of reagent is provided in the middle of the second supply pipe. Temperature of large amounts of reagents are brought close to isothermal temperature T ₂ in the thermal insulation cabinet.
JP 2001-275659 A

As described above, in order to approximate the temperature of the reagent thermostatic temperature T _2, it is necessary to increase the first supply pipe. When the first supply pipe is long, the amount of reagent remaining in the first supply pipe increases. That is, the amount of reagent supplied to the cell culture container varies. In addition, the reagent is wasted.

  Accordingly, the present invention has been made to solve the above-described problems, and provides a cell culture apparatus and a cell culture cassette that can reduce the residual amount of a reagent remaining in a supply pipe. Objective.

The cell culture device according to the first feature is a device for culturing cells. The cell culture apparatus includes a reagent container that stores a reagent at a cold temperature T ₁ , a cell culture container that stores cells cultured at a constant temperature T ₂ higher than the cold temperature T ₁ , the reagent container, and the cell culture A surface of the supply pipe between the reagent container and the cell culture container and a supply pipe constituting a flow path of a reagent supplied from the reagent container to the cell culture container. and a heating section for heating to a temperature T _3. The predetermined temperature _{T 3} is higher than the isothermal temperature _{T 2.}

According to this feature, between the reagent vessels and the cell culture vessel, the surface of the supply pipe is heated to a predetermined temperature T _3. The predetermined temperature _{T 3} is higher than the isothermal temperature _{T 2.}

Therefore, even if short supply pipe, it can be in the supply port to the cell culture vessel closer to the temperature of the reagent thermostatic temperature T _2. That is, the residual amount of the reagent remaining in the supply pipe can be reduced. This can suppress variations in the amount of reagent supplied to the cell culture container. In addition, the reagent can be used effectively.

  In the first feature, the reagent containers are a first reagent container for storing a first reagent and a second reagent container for storing a second reagent. The supply pipe is a first supply pipe that connects the first reagent container and the cell culture container, and a second supply pipe that connects the second reagent container and the cell culture container. The volume of the first supply pipe is smaller than the volume of the second supply pipe. The heating unit heats the first reagent flowing through the first supply pipe.

  According to this feature, the residual amount of the first reagent remaining in the first supply pipe can be reduced by heating the first reagent flowing through the first supply pipe having a volume smaller than that of the second supply pipe. . Thereby, variation in the amount of the first reagent supplied to the cell culture container can be suppressed. In addition, the first reagent can be used effectively.

  In the first feature, the cell culture device further includes an intermediate chamber for accommodating an intermediate pipe constituting a part of the first supply pipe between the reagent container and the cell culture container. The heating unit heats the first reagent flowing through the intermediate pipe in the intermediate chamber.

  According to this feature, since the heating unit heats the first reagent flowing through the intermediate pipe in the intermediate chamber, the influence of heating by the heating unit on the reagent container and the cell culture container provided outside the intermediate chamber is reduced. be able to.

In the first feature, the cell culture device further includes a refrigeration circuit including a compressor that compresses the refrigerant, a radiator that radiates the refrigerant, a decompressor that decompresses the refrigerant, and a heat absorber that absorbs heat from the refrigerant. The heat absorber may be used a reagent contained in the reagent container as a cooling unit for cooling the cold temperature T _1. The radiator is used as the heating unit.

  According to this feature, the radiator provided in the refrigeration circuit is used as a heating unit that heats the reagent flowing through the supply pipe. The heat absorber provided in the refrigeration circuit is used as a cooling unit for cooling the reagent accommodated in the reagent container. Therefore, since heat radiation (heating) and heat absorption (cooling) can be realized simultaneously by driving only the compressor, the power consumption of the cell culture device can be reduced. That is, the power consumption of the cell culture device can be reduced as compared with the case where the supply pipe is heated and the reagent is cooled by separate devices.

The cell culture cassette according to the second feature is provided in a cell culture apparatus for culturing cells. The cell culture cassette includes a reagent container that stores a reagent at a cold temperature T ₁ , a cell culture container that stores cells cultured at a constant temperature T ₂ higher than the cold temperature T ₁ , the reagent container, and the cell culture And a supply pipe that configures a flow path of a reagent supplied from the reagent container to the cell culture container. A part of the supply pipe constitutes an intermediate pipe between the reagent container and the cell culture container. Between said reagent container and the cell culture vessel, the surface of the intermediate pipe is heated to a predetermined temperature T _3. The predetermined temperature _{T 3} is higher than the isothermal temperature _{T 2.}

According to this feature, between the reagent vessels and the cell culture vessel, the surface of the intermediate pipe is heated to a predetermined temperature T _3. The predetermined temperature _{T 3} is higher than the isothermal temperature _{T 2.}

Therefore, even if short supply pipe, it can be in the supply port to the cell culture vessel closer to the temperature of the reagent thermostatic temperature T _2. That is, the residual amount of the reagent remaining in the supply pipe can be reduced. This can suppress variations in the amount of reagent supplied to the cell culture container. In addition, the reagent can be used effectively.

  2nd characteristic WHEREIN: The said reagent container is a 1st reagent container which accommodates a 1st reagent, and a 2nd reagent container which accommodates a 2nd reagent. The supply pipe is a first supply pipe that connects the first reagent container and the cell culture container, and a second supply pipe that connects the second reagent container and the cell culture container. The volume of the first supply pipe is smaller than the volume of the second supply pipe. The intermediate pipe constitutes a part of the first supply pipe.

  According to this feature, the residual amount of the first reagent remaining in the first supply pipe can be reduced by heating the first reagent flowing through the first supply pipe having a volume smaller than that of the second supply pipe. . Thereby, variation in the amount of the first reagent supplied to the cell culture container can be suppressed. In addition, the first reagent can be used effectively.

  ADVANTAGE OF THE INVENTION According to this invention, the cell culture apparatus and cell culture cassette which make it possible to reduce the residual amount of the reagent which remains in supply piping can be provided.

  Hereinafter, a cell culture device and a cell culture cassette according to an embodiment of the present invention will be described with reference to the drawings. In the following description of the drawings, the same or similar parts are denoted by the same or similar reference numerals.

  However, it should be noted that the drawings are schematic and ratios of dimensions are different from actual ones. Therefore, specific dimensions and the like should be determined in consideration of the following description. Moreover, it is a matter of course that portions having different dimensional relationships and ratios are included between the drawings.

[Outline of Embodiment]
The cell culture device according to the embodiment is a device for culturing cells. Cell culture device, a reagent container containing a reagent cold temperature T _1, and the cell culture vessel containing cells cultured at high isothermal temperature T ₂ than the cold temperature T _1, a reagent container and the cell culture vessel connected in which a supply pipe constituting the flow path of the reagent supplied from the reagent container to the cell culture vessel, between the reagent vessels and the cell culture vessel, heating unit for heating the surface of the supply pipe to the predetermined temperature T ₃ With. The predetermined temperature _{T 3} is higher than the isothermal temperature _{T 2.}

The cell culture cassette according to the embodiment is provided in a cell culture apparatus that cultures cells. Cell culture cassette, a reagent container containing a reagent cold temperature T _1, and the cell culture vessel containing cells cultured at high isothermal temperature T ₂ than the cold temperature T _1, a reagent container and the cell culture vessel And a supply pipe constituting a flow path of the reagent supplied from the reagent container to the cell culture container. A part of the supply pipe constitutes an intermediate pipe between the reagent container and the cell culture container. Between the reagent vessels and the cell culture vessel, the surface of the intermediate pipe is heated to a predetermined temperature T _3. The predetermined temperature _{T 3} is higher than the isothermal temperature _{T 2.}

Thus, in the embodiment, between the reagent vessels and the cell culture vessel, the surface of the supply pipe is heated to a predetermined temperature T _3. The predetermined temperature _{T 3} is higher than the isothermal temperature _{T 2.}

Therefore, even if short supply pipe, it can be in the supply port to the cell culture vessel closer to the temperature of the reagent thermostatic temperature T _2. That is, the residual amount of the reagent remaining in the supply pipe can be reduced. This can suppress variations in the amount of reagent supplied to the cell culture container. In addition, the reagent can be used effectively.

[First Embodiment]
(Configuration of cell culture device)
Hereinafter, the cell culture device according to the first embodiment will be described with reference to the drawings. FIG. 1 is a diagram showing a cell culture device 100 according to the first embodiment.

  As shown in FIG. 1, the cell culture device 100 includes a cold insulation zone 10, a heating zone 20, and a constant temperature zone 30. Specifically, the cell culture apparatus 100 includes a cell culture cassette 200, and the cell culture cassette 200 includes a cold insulation zone 10, a heating zone 20, and a constant temperature zone 30. In addition, the cell culture device 100 includes a cooling unit (cooling unit 11) and a heating unit (heating unit 21, heating unit 22, heating unit 31, heating unit 32).

The cold insulation zone 10 has a reagent container 210 that contains a reagent at a cold preservation temperature T ₁ (for example, 4 ° C.). The cooling unit 11 is a Peltier element or the like, and is provided outside the cold insulation zone 10. The cold insulation zone 10 is partitioned from the heating zone 20 and constitutes a cold insulation chamber.

The reagent accommodated in the reagent container 210 is cooled by the cooling unit 11 and is kept at the cold insulation temperature T ₁ by the cooling unit 11. The reagent is, for example, an enzyme, protein, inducer, introduction agent, serum, cytokine, medium, or phosphate buffered saline (PBS).

  The reagent accommodated in the reagent container 210 is supplied to a cell culture container 231 described later via a supply pipe 240. The supply pipe 240 connects the reagent container 210 and the cell culture container 231 and constitutes a flow path for the reagent supplied from the reagent container 210 to the cell culture container 231.

  The heating zone 20 accommodates an intermediate pipe 241 that constitutes a part of the supply pipe 240 between the cold insulation zone 10 (reagent container 210) and the constant temperature zone 30 (cell culture container 231). The heating unit 21 and the heating unit 22 are electric heaters or the like, and are provided outside the heating zone 20. The heating zone 20 is partitioned from the cold insulation zone 10 and the constant temperature zone 30 and constitutes an intermediate chamber.

  The intermediate pipe 241 is made of a metal material. The shape of the intermediate pipe 241 is a branched shape, a stitch shape, a flat shape, a single shape (one pipe), or the like.

The surface of the intermediate pipe is heated to a predetermined temperature T ₃ by the heating unit 21 and the heating portion 22. The predetermined temperature _{T 3} is isothermal temperature _{T 2} to be described later (for example, 37 ° C.) higher than. The predetermined temperature T ₃ is cold temperature T ₁ (e.g., 4 ° C.) is of course higher than.

For example, the predetermined temperature T ₃ is represented by a constant temperature T ₂ × α (> 1.0). α is, for example, 1.05. Here, the value of α is determined by the length of the intermediate pipe and the flow rate and flow rate of the reagent flowing through the intermediate pipe. For example, the shorter the intermediate pipe, the larger the value of α. The faster the reagent flowing through the intermediate pipe, the higher the value of α. The value of α increases as the flow rate of the reagent flowing through the intermediate pipe increases.

Incidentally, cold temperature T _1, the thermostatic temperature T ₂ and a predetermined temperature T ₃ is a control target value. Therefore, these actually measured values may have a certain range (for example, ± 0.25 ° C.) with respect to the control target value. Such a width depends on the performance of the cooling unit 11 (for example, a Peltier element), the heating unit 21, the heating unit 22, the heating unit 31, and the heating unit 32 (for example, an electric heater). Further, such a width, cold temperature T _1, dependent on the performance of the sensor for measuring the isothermal temperature T ₂ and a predetermined temperature T _3.

Here, the predetermined temperature T ₃ may be measured by a temperature sensor provided on the surface of the intermediate pipe. Heating unit 21 and the heating unit 22 is provided close to the surface of the intermediate pipe, it is possible to heat the surface of the intermediate pipe at a predetermined temperature T _3.

  The constant temperature zone 30 includes a cell culture vessel 231, a pump 232, a fan 233, and a humidifying pan 234. The heating unit 31 and the heating unit 32 are electric heaters or the like, and are provided outside the constant temperature zone 30. The constant temperature zone 30 is partitioned from the heating zone 20 and constitutes a constant temperature room.

The cell culture container 231 accommodates cells that are cultured at a constant temperature T ₂ (eg, 37 ° C.) that is higher than the cold temperature T ₁ . The contained accommodated into the cell culture vessel 231 (cells, medium and phosphate buffered solutions) is heated by the heating unit 31 and the heating unit 32 is kept at a constant temperature temperature T ₂ by the heating unit 31 and the heating portion 32.

  The pump 232 sends the reagent supplied from the reagent container 210 to the cell culture container 231 via the supply pipe 240. The fan 233 circulates air in the constant temperature zone 30. The humidifying pan 234 humidifies the inside of the constant temperature zone 30 and keeps the inside of the constant temperature zone 30 at approximately 100% humidity.

  Here, it should be noted that the cell culture cassette 200 is configured to be detachable from the cell culture apparatus 100. That is, the cell culture cassette 200 is configured to be replaceable.

(Function and effect)
In the first embodiment, between the reagent container 210 and the cell culture vessel 231, the surface of the supply pipe 240 (the intermediate pipe 241) is heated to a predetermined temperature _{T 3.} The predetermined temperature _{T 3} is higher than the isothermal temperature _{T 2.}

Therefore, even if short supply pipe 240, can be in the supply port to the cell culture vessel 231 close to the temperature of the reagent thermostatic temperature T _2. That is, the residual amount of the reagent remaining in the supply pipe 240 can be reduced. Thereby, variation in the amount of the reagent supplied to the cell culture container 231 can be suppressed. In addition, the reagent can be used effectively.

  For example, this is effective when an enzyme (such as trypsin) is used. In addition, when the configuration according to the first embodiment is applied to an expensive cytokine that is used in a small amount compared to an enzyme (such as trypsin), the residual amount of the cytokine is reduced, which is effective. That is, variations in the supply amount of cytokines that are used in small amounts can be suppressed, and expensive cytokines are not wasted.

  In the first embodiment, the heating unit 21 and the heating unit 22 heat the reagent flowing through the intermediate pipe 241 in the heating zone 20, so that the reagent container 210 and the cell culture container 231 provided outside the intermediate pipe 241 are heated. The influence which the heating by the part 21 and the heating part 22 gives can be reduced.

[Modification 1]
Hereinafter, Modification 1 of the first embodiment will be described with reference to FIG. In the following, differences from the first embodiment will be mainly described. In FIG. 2, it should be noted that the same components as those in FIG.

  Specifically, in the first modification, as illustrated in FIG. 2, the cooling unit 11 is provided not inside the cold insulation zone 10 but inside the cold insulation zone 10. The heating unit 21 and the heating unit 22 are provided not inside the heating zone 20 but inside the heating zone 20. The heating unit 31 is provided not inside the constant temperature zone 30 but inside the constant temperature zone 30.

  Thus, the heating unit 21 and the heating unit 22 directly heat the reagent flowing through the intermediate pipe 241 that constitutes a part of the supply pipe 240. The heating unit 31 directly heats the contents (cells, medium, and phosphate buffer solution) accommodated in the cell culture container 231.

  Thus, the heating zone 20 does not need to be partitioned from the constant temperature zone 30, and the heating zone 20 and the constant temperature zone 30 constitute one processing chamber.

[Modification 2]
Hereinafter, Modification 2 of the first embodiment will be described with reference to FIG. In the following, differences from the first embodiment will be mainly described. In FIG. 3, it should be noted that the same components as those in FIG.

  Specifically, in the modified example 2, as shown in FIG. 3, the cell culture device 100 includes a storage zone 40 in addition to the cold insulation zone 10, the heating zone 20, and the constant temperature zone 30. 3 is a schematic diagram of the cell culture device 100, and it should be noted that the above-described configuration is omitted in FIG.

  In addition, the cell culture device 100 includes a supply pipe 240IN that connects the reagent container 210 and the cell culture container 231 and a supply pipe 240OUT that connects the cell culture container 231 and the storage tank 41.

  Note that one end (injection port 245IN) of the supply pipe 240IN is preferably disposed so as to contact the wall surface of the cell culture vessel 231. As a result, the reagent is injected along the wall surface of the cell culture vessel 231, so that damage to the cells is reduced.

  Moreover, it is preferable that one end (discharge port 245OUT) of the supply pipe 240OUT is disposed in the vicinity of the bottom surface of the cell culture vessel 231. As a result, the cell suspension containing the cells cultured in the cell culture container 231 is efficiently discharged. That is, the cell suspension remaining in the cell culture vessel 231 is reduced.

  The cold insulation zone 10 includes a pump 12A, a pump 12B, a valve 13A, and a valve 13B. The pump 12A sends the reagent supplied from the reagent container 210 to the cell culture container 231 through the supply pipe 240IN. The pump 12B sends out the cells taken out from the cell culture container 231 to the storage tank 41 via the supply pipe 240OUT. The valve 13A adjusts the flow rate of the reagent flowing through the supply pipe 240IN. The valve 13B adjusts the flow rate of cells flowing through the supply pipe 240OUT.

  In the second modification, the cold insulation zone 10 is provided with a reagent kit 220 having a plurality of reagent containers 210 as a unit.

  The storage zone 40 is maintained at a storage temperature (for example, −18 ° C., −30 ° C., or −80 ° C.) and has a storage tank 41. The storage tank 41 stores the cells cultured with the medium in the cell culture container 231.

[Second Embodiment]
Hereinafter, the second embodiment will be described with reference to the drawings. In the following, differences from the first embodiment will be described.

  In the second embodiment, the reagent containers are a first reagent container that houses the first reagent and a second reagent container that houses the second reagent. The supply pipe is a first supply pipe that connects the first reagent container and the cell culture container, and a second supply pipe that connects the second reagent container and the cell culture container. The volume of the first supply pipe is smaller than the volume of the second supply pipe. The heating unit heats the first reagent flowing through the first supply pipe.

(Configuration of cell culture device)
Hereinafter, a cell culture device according to a second embodiment will be described with reference to the drawings. FIG. 4 is a diagram showing a cell culture device 100 according to the second embodiment. In FIG. 4, it should be noted that the same reference numerals are given to the same components as those in FIG.

  As shown in FIG. 4, the cold insulation zone 10 includes a first reagent container 210 </ b> A that contains the first reagent as a reagent container as a small amount of reagent (enzyme, protein, inducer, introduction agent, serum, cytokine, etc.), and a large amount. As a reagent (medium, PBS, etc.). It should be noted that the supply amount of the first reagent to the cell culture vessel 231 is smaller than the supply amount of the second reagent to the cell culture vessel 231.

  In addition, the cell culture apparatus 100 has a first supply pipe 240 that connects the first reagent container 210A and the cell culture container 231 and a second supply pipe 250 that connects the second reagent container 210B and the cell culture container 231 as supply pipes. And have. It should be noted that the volume of the first supply pipe 240 is smaller than the volume of the second supply pipe 250.

  Here, the first supply pipe 240 according to the second embodiment is the same as the supply pipe 240 according to the first embodiment. That is, a part of the first supply pipe 240 constitutes the intermediate pipe 241 according to the first embodiment.

The second supply pipe 250 has a sufficient length so that the temperature of the second reagent reaches the constant temperature T ₂ before the second reagent reaches the cell culture container 231. Specifically, an intermediate pipe 251 constituting a part of the second supply pipe 250 is provided in the constant temperature zone 30. The intermediate pipe 251 has, for example, a folded shape.

  In the second embodiment, the constant temperature zone 30 includes a plurality of pumps 232 (pumps 232A and 232B), a plurality of valves 235 (valves 235A, valves 235B and valves 235C), and a waste liquid tank 236.

  The pump 232 </ b> A sends the first reagent supplied from the first reagent container 210 </ b> A to the cell culture container 231 through the first supply pipe 240. The pump 232 </ b> B sends the second reagent supplied from the second reagent container 210 </ b> B to the cell culture container 231 through the second supply pipe 250.

  The valve 235 </ b> A adjusts the flow rate of the first reagent flowing through the first supply pipe 240. The valve 235B adjusts the flow rate of the second reagent flowing through the second supply pipe 250. The valve 235 </ b> C adjusts the flow rate of the waste liquid flowing through the pipe connecting the cell culture container 231 and the waste liquid tank 236.

  The waste liquid tank 236 stores a mixed liquid (waste liquid) such as a culture medium and a phosphate buffer solution.

  In the second embodiment, the cell culture device 100 includes a drying chamber 260. The drying chamber 260 has a communication path 261 that allows the heating zone 20 and the constant temperature zone 30 to communicate with each other. A part of the communication path 261 is provided adjacent to the cold insulation zone 10.

Thereby, the humid gas flowing from the constant temperature zone 30 is condensed in a part of the communication path 261 provided adjacent to the cold insulation zone 10, and the gas flowing through the communication path 261 is dried. This can gas warmed in a constant temperature zone 30 is used for the temperature rise of the heating zone 20, raising the temperature of the heating zone 20 to efficiently predetermined temperature T _3.

  Moreover, since the drying chamber 260 dries the humid gas flowing from the constant temperature zone 30, it is possible to suppress the occurrence of condensation in the heating zone 20.

(Cell culture method)
Hereinafter, the cell culture method according to the second embodiment will be described with reference to the drawings. 5 and 6 are flowcharts showing a cell culture method according to the second embodiment.

  As a premise of the processing shown in FIGS. 5 and 6, the cold insulation zone 10 is cooled by the cooling unit 11, and the constant temperature zone 30 is heated by the heating unit 31 and the heating unit 32.

  As shown in FIG. 5, in step 10, various settings are performed. Various settings are performed in a sterile environment using a safety cabinet or an isolator. For example, the first reagent container 210A, the second reagent container 210B, and the cell culture container 231 are set.

  In step 20, the cell culture device 100 turns on the heating unit 21 that heats the heating zone 20.

  In step 30, the cell culture device 100 opens the valve 235A and the valve 235B.

  In step 40, the cell culture device 100 injects the reagent into the cell culture container 231. Specifically, the cell culture device 100 injects the first reagent into the cell culture container 231 via the first supply pipe 240. The cell culture device 100 injects the second reagent into the cell culture container 231 via the second supply pipe 250.

  In step 50, the cell culture device 100 turns off the heating unit 21 that heats the heating zone 20.

  In step 60, the cell culture device 100 closes the valve 235A and the valve 235B.

  In step 70, the cell culture device 100 starts culturing cells in the cell culture container 231.

  In step 80, the cell culture device 100 interrupts the cell culture and restarts the cell culture. Details of step 80 will be described later (see FIG. 6).

  In step 90, the cell culture device 100 ends the cell culture in the cell culture container 231.

  In step 100, the cell culture container 231 is taken out, and the cultured cells are taken out from the cell culture container 231.

  Next, details of the culture interruption / resumption process (step 80) will be described with reference to FIG.

  As shown in FIG. 6, in step 81, the cell culture device 100 opens the valve 235 </ b> C and discharges the medium stored in the cell culture container 231 to the waste liquid tank 236.

  In step 82, the cell culture device 100 opens the valve 235 </ b> B and injects the second reagent (medium) into the cell culture container 231 through the second supply pipe 250.

  In step 83, the cell culture device 100 turns on the heating unit 21 that heats the heating zone 20.

  In step 84, the cell culture device 100 opens the valve 235A.

  In step 85, the cell culture device 100 injects the first reagent into the cell culture container 231 via the first supply pipe 240.

  In step 86, the cell culture device 100 turns off the heating unit 21 that heats the heating zone 20.

  In step 87, the cell culture device 100 closes the valve 235A.

  In step 88, the cell culture device 100 resumes cell culture in the cell culture container 231.

(Function and effect)
In the second embodiment, the same effect as in the first embodiment can be obtained for the first reagent with a small amount supplied to the cell culture container 231. That is, the residual amount of the first reagent remaining in the first supply pipe 240 can be reduced. Thereby, variation in the amount of the first reagent supplied to the cell culture container 231 can be suppressed. In addition, the first reagent can be used effectively.

[Third Embodiment]
Hereinafter, a third embodiment of the first embodiment will be described with reference to the drawings. In the following, differences from the first embodiment will be mainly described.

Specifically, in the third embodiment, the cell culture device 100 further includes a refrigeration circuit having a compressor that compresses the refrigerant, a radiator that dissipates the refrigerant, a decompressor that decompresses the refrigerant, and a heat absorber that absorbs heat from the refrigerant. Prepare. Heat sink is used as a cooling unit for cooling the reagent contained in the reagent container cold temperature T _1. The radiator is used as a heating unit that heats the reagent flowing through the supply pipe (intermediate pipe) between the reagent container and the cell culture container.

(Configuration of cell culture device)
Hereinafter, a cell culture device according to a third embodiment will be described with reference to the drawings. FIG. 7 is a diagram showing a cell culture device 100 according to the third embodiment. In FIG. 7, it should be noted that the same components as those in FIG.

  As shown in FIG. 7, the cell culture device 100 includes a refrigeration circuit 300. The refrigeration circuit 300 includes a heat absorber 310, a compressor 320, a radiator 330, a decompressor 340, and a refrigerant channel 350.

  Here, an HFC (Hydro Fluoro Carbon) refrigerant or an HC (Hydro Carbon) refrigerant will be described as an example of the refrigerant circulating in the refrigerant flow path 350. Further, the circulation of the refrigerant will be described with reference to FIG. In FIG. 8, the vertical axis represents the pressure (P) with respect to the refrigerant, and the horizontal axis represents the enthalpy (h) of the refrigerant. The isotherm is a line indicating a combination of pressure (P) and enthalpy (h) at which the temperature is constant. The saturated liquid line is a line indicating the boundary between the supercooled liquid and the wet steam, and the saturated vapor line is a line indicating the boundary between the wet steam and the superheated steam. The critical point is the boundary between the saturated liquid line and the saturated vapor line.

  The heat absorber 310 absorbs heat by the refrigerant circulating in the refrigerant flow path 350. Specifically, the heat absorber 310 is used as a cooling unit that cools the reagent stored in the reagent container 210. In FIG. 8, as shown in step (1), the enthalpy (h) increases due to heat absorption by the refrigerant while the pressure (P) remains constant.

  The compressor 320 compresses the refrigerant evaporated in the heat absorber 310. In FIG. 8, as shown in step (2), the degree of superheat of the refrigerant increases as the pressure (P) increases.

  The radiator 330 radiates the heat of the refrigerant compressed by the compressor 320. Specifically, the radiator 330 is used as a heating unit that heats the reagent flowing through the supply pipe 240 (intermediate pipe 241).

  Specifically, a part 331 of the radiator 330 provided on the refrigerant channel 350 on the side close to the compressor 320 is used as a heating unit that heats the reagent flowing through the supply pipe 240 (intermediate pipe 241). On the other hand, a part 332 of the radiator 330 provided on the refrigerant flow path 350 on the side close to the decompressor 340 is used as a heating unit that heats cells cultured in the cell culture vessel 231.

  In FIG. 8, as shown in step (3), the enthalpy (h) decreases while the pressure (P) remains constant due to the cooling of the refrigerant. As a result, the refrigerant transitions to the supercooled liquid.

  The decompressor 340 decompresses the refrigerant radiated by the radiator 330. In FIG. 8, as shown in step (4), the pressure (P) decreases while the enthalpy (h) remains constant. As a result, the refrigerant transitions to wet steam.

  The refrigerant channel 350 is a refrigerant channel. Specifically, the refrigerant channel 350 is an annular channel that passes through the heat absorber 310, the compressor 320, the radiator 330, and the decompressor 340.

(Function and effect)
In 3rd Embodiment, the heat radiator 330 provided in the refrigerating circuit 300 is used as a heating part which heats the reagent which flows through the supply piping 240 (intermediate piping 241). The heat absorber 310 provided in the refrigeration circuit 300 is used as a cooling unit that cools the reagent accommodated in the reagent container 210. Therefore, since heat dissipation (heating) and heat absorption (cooling) can be realized simultaneously by driving only the compressor 320, the power consumption of the cell culture device 100 can be reduced. That is, the power consumption of the cell culture device 100 can be reduced as compared to the case where the supply pipe 240 is heated and the reagent is cooled by separate devices.

[Other Embodiments]
Although the present invention has been described with reference to the above-described embodiments, it should not be understood that the descriptions and drawings constituting a part of this disclosure limit the present invention. From this disclosure, various alternative embodiments, examples and operational techniques will be apparent to those skilled in the art.

  For example, in the third embodiment, a vibration isolation structure may be provided in the refrigeration circuit 300 so that the vibration of the compressor 320 is not transmitted to the cell culture container 231. The vibration isolation structure is, for example, a vibration isolation member or a spring suspension.

It is a figure showing cell culture device 100 concerning a 1st embodiment. It is a figure which shows the cell culture apparatus 100 which concerns on the example 1 of a change. It is a figure which shows the cell culture apparatus 100 which concerns on the example 2 of a change. It is a figure which shows the cell culture apparatus 100 which concerns on 2nd Embodiment. It is a flowchart which shows the cell culture method which concerns on 2nd Embodiment. It is a flowchart which shows the cell culture method which concerns on 2nd Embodiment. It is a figure which shows the cell culture apparatus 100 which concerns on 3rd Embodiment. It is a figure for demonstrating the refrigerant | coolant which concerns on 3rd Embodiment.

Explanation of symbols

  DESCRIPTION OF SYMBOLS 10 ... Cold storage zone, 11 ... Cooling part, 12 ... Pump, 13 ... Valve, 20 ... Heating zone, 21, 22 ... Heating part, 30 ... Constant temperature zone, 31, 32 ... Heating part, 40 ... Storage zone, 41 ... Storage Tank, 100 ... Cell culture device, 200 ... Cell culture cassette, 210 ... Reagent container, 211 ... Reagent kit, 231 ... Cell culture container, 232 ... Pump, 233 ... Fan, 234 ... Humidification pan, 235 ... Valve, 236 ... Waste liquid Tank, 240 ... supply pipe (first supply pipe), 241 ... intermediate pipe, 250 ... second supply pipe, 251 ... intermediate pipe, 260 ... drying chamber, 261 ... communication path, 300 ... refrigeration circuit, 310 ... heat absorber, 320 ... Compressor, 330 ... Radiator, 340 ... Pressure reducer, 350 ... Refrigerant flow path

Claims (6)

A cell culture device for culturing cells,
A reagent container for storing the reagent at a cold temperature T ₁ ;
A cell culture container for accommodating cells cultured at a constant temperature T ₂ higher than the cold-retaining temperature T ₁ ;
A supply pipe that connects the reagent container and the cell culture container, and constitutes a flow path of a reagent that is supplied from the reagent container to the cell culture container;
Between said reagent container and the cell culture vessel, and a heating unit for heating the surface of the supply pipe to the predetermined temperature T _3,
The predetermined temperature T _3, the cell culture apparatus characterized by higher than the isothermal temperature T _2. The reagent containers are a first reagent container for storing a first reagent and a second reagent container for storing a second reagent,
The supply pipe is a first supply pipe that connects the first reagent container and the cell culture container, and a second supply pipe that connects the second reagent container and the cell culture container,
The volume of the first supply pipe is smaller than the volume of the second supply pipe,
The cell culture device according to claim 1, wherein the heating unit heats the first reagent flowing through the first supply pipe. Further comprising an intermediate chamber for accommodating an intermediate pipe constituting a part of the first supply pipe between the reagent container and the cell culture container,
The cell culture device according to claim 2, wherein the heating unit heats the first reagent flowing through the intermediate pipe in the intermediate chamber. A compressor that compresses the refrigerant; a radiator that dissipates the refrigerant; a decompressor that decompresses the refrigerant; and a refrigeration circuit that absorbs heat from the refrigerant;
The heat absorber may be used a reagent contained in the reagent container as a cooling unit for cooling the cold temperature T _1,
The cell culture device according to claim 1, wherein the radiator is used as the heating unit. A cell culture cassette provided in a cell culture device for culturing cells,
A reagent container for storing the reagent at a cold temperature T ₁ ;
A cell culture container for accommodating cells cultured at a constant temperature T ₂ higher than the cold-retaining temperature T ₁ ;
Connecting the reagent container and the cell culture container, and comprising a supply pipe constituting a flow path of a reagent supplied from the reagent container to the cell culture container,
A part of the supply pipe constitutes an intermediate pipe between the reagent container and the cell culture container,
Between said reagent container and the cell culture vessel, the surface of the intermediate pipe is heated to a predetermined temperature T _3,
The predetermined temperature T _3, the cell culture cassette being higher than the isothermal temperature T _2. The reagent containers are a first reagent container for storing a first reagent and a second reagent container for storing a second reagent,
The supply pipe is a first supply pipe that connects the first reagent container and the cell culture container, and a second supply pipe that connects the second reagent container and the cell culture container,
The volume of the first supply pipe is smaller than the volume of the second supply pipe,
The cell culture cassette according to claim 5, wherein the intermediate pipe constitutes a part of the first supply pipe.

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