Patent Application
20210102158
The technique disclosed relates to a cell culture apparatus and a cell culture method.
The following techniques are known as techniques relating to a cell culture apparatus. For example, JP2015-503361A describes a heat exchange module used in a chemical, pharmaceutical or biological reactor system. The heat exchange module is configured to be disposed in a reactor system having a flexible single-use container and includes at least one heat conductive surface that is made in contact with the flexible single-use container to promote heat transfer and a fluid circulation flow path through which a heat exchange fluid can be circulated.
In cell culture, a cell suspension containing cells and a medium is stored in an incubator maintained at a predetermined temperature (for example, 37° C.) suitable for cell growth, in a state of being contained in a culture vessel. In a case of performing a necessary treatment such as a medium exchange treatment and a subculture treatment on the cell suspension, the cell suspension is transferred to an outside of the incubator and the necessary treatment is carried out in a room temperature environment. In this case, the temperature of the cell suspension is lowered to a temperature lower than the predetermined temperature suitable for cell growth. After the necessary treatment is completed, the cell suspension is contained in the incubator again. Therefore, the temperature of the cell suspension rises to the predetermined temperature suitable for cell growth. In a case where the temperature rising rate of the cell suspension in this case is low, the cells are exposed to a low temperature for a long time, which may adversely affect growing property and viability of the cells. This problem becomes more remarkable in a case where large-scale culture is carried out. That is, in the large-scale culture, a heat capacity of a cell suspension, which is an object of temperature rising, becomes larger, comparing to a case of small-scale culture, and the time during which the cells are exposed to the low temperature is prolonged. In the large-scale culture, one of reasons that the time during which the cells are exposed to the low temperature is prolonged is that the time required for a medium exchange treatment and a subculture treatment becomes long in the large-scale culture.
Moreover, in a general incubator, an atmospheric temperature inside the incubator is likely to be uneven due to the influence of a position of a heat source, a disposition of a blower fan, a direction of an air blow, a position of a heat insulating material inside the incubator, and a position of a door. Particularly in the large-scale culture in which a cell suspension is contained in a large-sized culture vessel to culture cells, in a case where the atmospheric temperature inside the incubator is uneven, a temperature of the cell suspension in the culture vessel becomes nonuniform. As a result, qualities of cells become nonuniform, and there is a concern that culture efficiency may decrease. In addition, in a general incubator, since a temperature of the cell suspension which is a temperature rising target is raised by heat conduction using an atmosphere inside the incubator having a large specific heat as a heat transfer medium, the temperature unevenness generated in the cell suspension is not easily eliminated.
The disclosed technique has been made in view of the above points, and an object thereof is to improve a temperature rising rate of a temperature rising target and enhance a temperature uniformity of the temperature rising target.
A cell culture apparatus according to the disclosed technique includes: a heat transfer medium having a recess part or a plurality of hole parts in a contact surface with a culture vessel for containing a cell suspension; and an air supply port which communicates with the recess part or each of the plurality of hole parts and to which a gas is supplied.
A cell culture method according to the disclosed technique includes: bringing a culture vessel, in which a cell suspension is contained and which is gas-permeable, into contact with a surface of a heat transfer medium which is heated and has a recess part or a plurality of hole parts on the surface; and allowing a gas to flow through a gap which is formed by the surface of the heat transfer medium being brought into contact with the culture vessel.
Hereinafter, embodiments of the disclosed technique will be described with reference to the drawings. In the drawings, substantially the same or equivalent constituent elements or parts are designated by the same reference numerals.
The cell culture apparatus 1 includes a heat transfer medium 10 that is in contact with a surface of the culture vessel 50. The heat transfer medium 10 includes a first part 11 disposed on one surface side of the culture vessel 50 and a second part 12 disposed on the other surface side of the culture vessel 50. When using the cell culture apparatus 1, the culture vessel 50 is sandwiched between the first part 11 and the second part 12 of the heat transfer medium 10. The heat transfer medium 10 is made of a material having a relatively high thermal conductivity, such as metal and resin.
Each of the first part 11 and the second part 12 of the heat transfer medium 10 has a plurality of hole parts 20 on a contact surface S1 in contact with the culture vessel 50.
An air supply port 31a is provided at one end part of each of the first part 11 and the second part 12 of the heat transfer medium 10. The air supply port 31a is connected to the communication passage 30 via an air supply passage 31 provided inside the heat transfer medium 10. In addition, an exhaust port 32a is provided at the other end part of each of the first part 11 and the second part 12 of the heat transfer medium 10. The exhaust port 32a is connected to the communication passage 30 via an exhaust passage 32 provided inside the heat transfer medium 10. That is, each of the plurality of hole parts 20 communicates with both the air supply port 31a and the exhaust port 32a.
The plurality of hole parts 20 may be formed, for example, by machining a base material forming the heat transfer medium 10. In this case, it is preferable that the plurality of hole parts 20 is uniformly and regularly disposed on the contact surface S1 of the heat transfer medium 10, and it is preferable that a part of the heat transfer medium 10 in contact with the culture vessel 50 (a part other than the hole parts 20) extends uniformly on the contact surface S1 of the heat transfer medium 10. In addition, as the heat transfer medium 10, a porous material such as a metal sintered body may be used. In this case, a plurality of voids formed in the porous material function as the hole parts 20. In addition, as the heat transfer medium 10, an aggregate of a plurality of fibrous members, such as steel wool may be used. In this case, a plurality of voids formed between the fibrous members function as the hole parts 20.
The heat transfer medium 10 is configured such that a distance between the first part 11 and the second part 12 maintains a constant distance according to the thickness of the culture vessel 50. Accordingly, a deformation of the culture vessel 50 sandwiched between the first part 11 and the second part 12 is regulated.
Hereinafter, an example of a cell culture method using the cell culture apparatus 1 will be described. First, the culture vessel 50 is disposed between the first part 11 and the second part 12 of the heat transfer medium 10. The culture vessel 50 is disposed so that the gas flow port 51 is positioned vertically upward and the liquid flow port 52 is positioned vertically downward.
Next, the cell suspension 60 containing a plurality of cells to be cultured and a medium is allowed to flow into the culture vessel 50 through the liquid flow port 52. Thereafter, a gas is allowed to flow into the culture vessel 50 through the gas flow port 51. It is preferable that pressure inside the culture vessel 50 is kept at a state of the same as or higher than the pressure of an atmosphere outside the culture vessel 50. Accordingly, the culture vessel 50 is kept in close contact with the respective contact surfaces S1 of the first part 11 and the second part 12 of the heat transfer medium 10. A deformation of the culture vessel 50 due to the inflow of the cell suspension 60 into the culture vessel 50 is regulated by sandwiching the culture vessel 50 between the first part 11 and the second part 12. Thus, the thickness of the culture vessel 50 containing the cell suspension 60 is substantially uniform in the entire vertical direction.
Next, the temperature-adjusted and composition-adjusted gas is introduced into the heat transfer medium 10 through the respective air supply ports 31a of the first part 11 and the second part 12 of the heat transfer medium 10. The temperature of the gas introduced into the heat transfer medium 10 is adjusted to a temperature (for example, 37° C.) suitable for promoting the growth of cells contained in the culture vessel 50, for example. Further, the gas introduced into the heat transfer medium 10 contains, for example, oxygen and carbon dioxide necessary for promoting the growth of the cells contained in the culture vessel 50, and a mixing ratio thereof is adjusted. The gas introduced into the heat transfer medium 10 may contain water vapor.
The gas introduced from the air supply port 31a spreads throughout the heat transfer medium 10 via the air supply passage 31 and the communication passage 30. A part of the gas is discharged from each of the hole parts 20, and a part of the remaining gas is discharged from the exhaust port 32a via the exhaust passage 32. The temperature-adjusted gas spreads throughout the heat transfer medium 10, so that the temperature of the heat transfer medium 10 quickly rises to a desired temperature (for example, 37° C.). Since the contact surfaces S1 of the first part 11 and the second part 12 of the heat transfer medium 10 are in contact with the culture vessel 50, the temperature of the cell suspension 60 contained in the culture vessel 50 quickly rises to the desired temperature (for example, 37° C.). In addition, since the part of the heat transfer medium 10 in contact with the culture vessel 50 (the part other than the hole parts 20) extends uniformly over the entire contact surface S1, occurrence of temperature unevenness of the cell suspension 60 contained in the culture vessel 50 is suppressed and it is possible to enhance the temperature uniformity of the cell suspension 60.
Further, the temperature-adjusted and composition-adjusted gas discharged from the plurality of hole parts 20 provided in the contact surface S1 of the heat transfer medium 10 is supplied to the surface of the culture vessel 50 which is gas-permeable, permeates into the culture vessel 50, and is taken into the cell suspension 60. The composition-adjusted gas is taken into the cell suspension 60 to promote the growth of the cells.
In a case where the cell suspension 60 contained in the culture vessel 50 is allowed to flow out of the liquid flow port 52, the gas is introduced into the culture vessel 50 to pressurize the inside of the culture vessel 50, for example. Expansion of the culture vessel 50 due to the pressurization inside the culture vessel 50 is suppressed by sandwiching the culture vessel 50 between the first part 11 and the second part 12 of the heat transfer medium 10. Accordingly, it becomes easy to control the pressure inside the culture vessel 50, and it becomes easy to control the outflow amount of the cell suspension 60 from the culture vessel 50.
As described above, the cell culture apparatus 1 according to the embodiment of the disclosed technique includes the heat transfer medium 10 having the plurality of hole parts 20 on the contact surface S1 with the culture vessel 50, and allows the temperature-adjusted and composition-adjusted gas to flow through the gap which is formed by the contact surface S1 of the heat transfer medium 10 being brought into contact with the culture vessel 50. The heat of temperature-adjusted gas is conducted to the cell suspension 60 contained in the culture vessel 50 via the heat transfer medium 10, and the temperature of the cell suspension 60 rises to a desired temperature.
Since the heat transfer medium 10 formed of a solid material has a smaller specific heat than that of the gas, the temperature rising rate of the cell suspension 60 can be enhanced by raising the temperature of the cell suspension 60 via the heat transfer medium 10, comparing to a case of using a general incubator. Accordingly, it possible to suppress the time during which the cells are exposed to the low temperature from being prolonged, and it is possible to improve proliferation and a survival rate of the cells.
In addition, a part of the heat transfer medium 10 in contact with the culture vessel 50 (the part other than the hole parts 20) extends uniformly over the entire contact surface S1. In other words, the heat transfer medium 10 is in contact with the culture vessel 50 without deviation. Accordingly, occurrence of temperature unevenness of the cell suspension 60 contained in the culture vessel 50 is suppressed and it is possible to enhance the temperature uniformity of the cell suspension 60.
As described above, according to the cell culture apparatus 1 according to the embodiment of the disclosed technique, it is possible to improve the temperature rising rate of a temperature rising target and enhance the temperature uniformity of the temperature rising target. In addition, it is possible to carry out a temperature control of the cell suspension 60 and gas supply to the cell suspension 60 in parallel. The cell culture apparatus 1 according to an embodiment of the disclosed technique has a remarkable effect, particularly in a case of being applied to a large-scale culture in which a cell suspension is contained in a large-sized culture vessel to culture cells.
In addition, the expansion of the culture vessel 50 due to the pressurization inside the culture vessel 50 is suppressed by sandwiching the culture vessel 50 between the first part 11 and the second part 12 of the heat transfer medium 10. Accordingly, it becomes easy to control the pressure inside the culture vessel 50, and it becomes easy to control the outflow amount of the cell suspension 60 from the culture vessel 50.
The heater 40 is adjusted to a temperature (for example, 37° C.) suitable for promoting the growth of cells contained in the culture vessel 50. The heat generated from the heater 40 is conducted to the cell suspension 60 contained in the culture vessel 50 via the heat transfer medium 10, and the temperature of the cell suspension 60 rises to a desired temperature. It is noted that, in order to heat the heat transfer medium 10, a temperature-adjusted gas may be introduced into the heat transfer medium 10 from the air supply port 31a.
According to the cell culture apparatus 1A according to the present embodiment, as in the cell culture apparatus 1 according to the first embodiment, it is possible to improve the temperature rising rate of a temperature rising target and enhance the temperature uniformity of the temperature rising target. In addition, it is possible to carry out a temperature control of the cell suspension 60 and gas supply to the cell suspension 60 in parallel. In addition, according to the cell culture apparatus 1A according to the present embodiment, the heat transfer medium 10 is heated by the heater 40. Therefore, the temperature rising rate of the heat transfer medium 10 can be enhanced, comparing to a case where the heat transfer medium 10 is heated by only the temperature-adjusted gas.
Each of the first part 11 and the second part 12 of the heat transfer medium 10 has a recess part 21 and a protrusion part 22 on the contact surface S1 in contact with the culture vessel 50.
As shown in
An uneven structure including the recess part 21 and the protrusion parts 22 formed on the contact surface S1 of the heat transfer medium 10 with the culture vessel 50 may be formed by, for example, machining a base material forming the heat transfer medium 10. In this case, it is preferable that the recess part 21 and the protrusion parts 22 is uniformly and regularly disposed on the contact surface S1 of the heat transfer medium 10, and it is preferable that parts of the heat transfer medium 10 in contact with the culture vessel 50 (that is, the protrusion part 22) extend uniformly on the contact surface S1 of the heat transfer medium 10. The heat transfer medium 10 is configured such that a distance between the first part 11 and the second part 12 maintains a constant distance according to the thickness of the culture vessel 50.
Hereinafter, an example of a cell culture method using the cell culture apparatus 1B will be described. First, the culture vessel 50 is disposed between the first part 11 and the second part 12 of the heat transfer medium 10. The culture vessel 50 is disposed so that the gas flow port 51 is positioned vertically upward and the liquid flow port 52 is positioned vertically downward.
Next, the cell suspension 60 containing a plurality of cells to be cultured and a medium is allowed to flow into the culture vessel 50 through the liquid flow port 52. Thereafter, a gas is allowed to flow into the culture vessel 50 through the gas flow port 51. It is preferable that pressure inside the culture vessel 50 is kept at a state of the same as or higher than the pressure of an atmosphere outside the culture vessel 50. Accordingly, the culture vessel 50 is kept in close contact with the respective contact surfaces S1 of the first part 11 and the second part 12 of the heat transfer medium 10. A deformation of the culture vessel 50 due to the inflow of the cell suspension 60 into the culture vessel 50 is regulated by sandwiching the culture vessel 50 between the first part 11 and the second part 12. Thus, the thickness of the culture vessel 50 containing the cell suspension 60 is substantially uniform in the entire vertical direction.
Next, the temperature-adjusted and composition-adjusted gas is introduced into the heat transfer medium 10 through the respective air supply ports 31a of the first part 11 and the second part 12 of the heat transfer medium 10. The temperature of the gas introduced into the heat transfer medium 10 is adjusted to a temperature (for example, 37° C.) suitable for promoting the growth of cells contained in the culture vessel 50, for example. Further, the gas introduced into the heat transfer medium 10 contains, for example, oxygen and carbon dioxide necessary for promoting the growth of the cells contained in the culture vessel 50, and a mixing ratio thereof is adjusted. The gas introduced into the heat transfer medium 10 may contain water vapor.
The gas introduced from the air supply port 31a is supplied to the recess part 21 formed on the contact surface S1 of the heat transfer medium 10 via the air supply passage 31, flows along the recess part 21, spreads throughout the heat transfer medium 10, and is discharged from the exhaust port 32a via the exhaust passage 32.
The temperature-adjusted gas spreads throughout the heat transfer medium 10, so that the temperature of the heat transfer medium 10 quickly rises to a desired temperature (for example, 37° C.). Since the contact surfaces S1 of the first part 11 and the second part 12 of the heat transfer medium 10 are in contact with the culture vessel 50, the temperature of the cell suspension 60 contained in the culture vessel 50 quickly rises to the desired temperature (for example, 37° C.). In addition, the recess part 21 and the protrusion parts 22 are uniformity and regularly disposed on the contact surface S1 of the heat transfer medium 10, and parts of the heat transfer medium 10 in contact with the culture vessel 50 (that is, protrusion parts 22) extend uniformly on the contact surface S1 of the heat transfer medium 10. Therefore, occurrence of temperature unevenness of the cell suspension 60 contained in the culture vessel 50 is suppressed and it is possible to enhance the temperature uniformity of the cell suspension 60.
Further, the gas flowing along the recess part 21 provided in the contact surface S1 of the heat transfer medium 10 is supplied to the surface of the culture vessel 50 which is gas-permeable, permeates into the culture vessel 50, and is taken into the cell suspension 60. The composition-adjusted gas is taken into the cell suspension 60 to promote the growth of the cells.
In a case where the cell suspension 60 contained in the culture vessel 50 is allowed to flow out of the liquid flow port 52, the gas is introduced into the culture vessel 50 through the gas flow port 51 to pressurize the inside of the culture vessel 50, for example. Expansion of the culture vessel 50 due to the pressurization inside the culture vessel 50 is suppressed by sandwiching the culture vessel 50 between the first part 11 and the second part 12 of the heat transfer medium 10. Accordingly, it becomes easy to control the pressure inside the culture vessel 50, and it becomes easy to control the outflow amount of the cell suspension 60 from the culture vessel 50.
As described above, the cell culture apparatus 1B according to the present embodiment includes the heat transfer medium 10 having the recess part 21 and the protrusion parts 22 on the contact surface S1 with the culture vessel 50, and allows the temperature-adjusted and composition-adjusted gas to flow through the gap which is formed by the contact surface S1 of the heat transfer medium 10 being brought into contact with the culture vessel 50. The heat of temperature-adjusted gas is conducted to the cell suspension 60 contained in the culture vessel 50 via the heat transfer medium 10, and the temperature of the cell suspension 60 rises to a desired temperature.
According to the cell culture apparatus 1B according to the present embodiment, as in the cell culture apparatus 1 according to the first embodiment, it is possible to improve the temperature rising rate of a temperature rising target and enhance the temperature uniformity of the temperature rising target. In addition, it is possible to carry out a temperature control of the cell suspension 60 and gas supply to the cell suspension 60 in parallel.
In addition, the expansion of the culture vessel 50 due to the pressurization inside the culture vessel 50 is suppressed by sandwiching the culture vessel 50 between the first part 11 and the second part 12 of the heat transfer medium 10. Accordingly, it becomes easy to control the pressure inside the culture vessel 50, and it becomes easy to control the outflow amount of the cell suspension 60 from the culture vessel 50.
The heater 40 is adjusted to a temperature (for example, 37° C.) suitable for promoting the growth of cells contained in the culture vessel 50. The heat generated from the heater 40 is conducted to the cell suspension 60 contained in the culture vessel 50 via the heat transfer medium 10, and the temperature of the cell suspension 60 rises to a desired temperature. It is noted that, in order to heat the heat transfer medium 10, a temperature-adjusted gas may be introduced into the heat transfer medium 10 from the air supply port 31a.
According to the cell culture apparatus 1C according to the present embodiment, as in the cell culture apparatus 1 according to the first embodiment, it is possible to improve the temperature rising rate of a temperature rising target and enhance the temperature uniformity of the temperature rising target. In addition, it is possible to carry out a temperature control of the cell suspension 60 and gas supply to the cell suspension 60 in parallel. In addition, according to the cell culture apparatus 1C according to the present embodiment, the heat transfer medium 10 is heated by the heater 40. Therefore, the temperature rising rate of the heat transfer medium 10 can be enhanced, comparing to a case where the heat transfer medium 10 is heated by only the temperature-adjusted gas.
According to the cell culture system 100 of the present embodiment, it is possible to suppress a temperature change inside the culture vessel 50 and a composition change of the gas introduced into the culture vessel 50.
As shown in
In the present embodiment, the case where one or more cell culture apparatuses 1 are housed in the housing container 70 is illustrated. However, it is also possible to adopt the cell culture apparatus 1A (
According to the cell culture apparatus of the present disclosure, it is possible to improve a temperature rising rate of a cell suspension which is a temperature rising target and enhance a temperature uniformity of the cell suspension.
It is preferable that the recess part or the plurality of hole parts is disposed uniformly over an entire contact surface of the heat transfer medium.
Accordingly, it possible to promote an effect of increasing the temperature uniformity of the cell suspension.
The heat transfer medium may include a porous material. In addition, the contact surface of the heat transfer medium may have an uneven structure including the recess part and a protrusion part.
It is preferable that a part of the heat transfer medium in contact with the culture vessel extends uniformly on the contact surface of the heat transfer medium.
Accordingly, it possible to promote an effect of increasing the temperature uniformity of the cell suspension.
The heat transfer medium may include a first part provided on one surface side of the culture vessel and a second part provided on the other surface side of the culture vessel opposite to the one surface.
Accordingly, expansion of the culture vessel due to pressurization inside the culture vessel is suppressed, and it becomes easy to control pressure inside the culture vessel. As a result, it becomes easy to control the outflow amount of the cell suspension from the culture vessel.
The cell culture apparatus according to the disclosed technique may further include a heater attached to the heat transfer medium.
As a result, it is possible to increase the temperature rising rate of the heat transfer medium, comparing to a case where the heat transfer medium is heated only by a temperature-adjusted gas.
It is preferable that a width of the recess part is at most 2.5 times a thickness of a material forming the culture vessel, and a depth of the recess part is at least 0.8 times the thickness of the material forming the culture vessel.
Accordingly, it is possible to prevent a material forming the culture vessel from closing the recess part.
According to the cell culture method of the present disclosure, it is possible to improve a temperature rising rate of a cell suspension which is a temperature rising target and enhance a temperature uniformity of the cell suspension.
The heat transfer medium may be heated by using a temperature-adjusted gas as the gas allowed to flow through a gap which is formed by the contact surface of the heat transfer medium being brought into contact with the culture vessel. In addition, the heat transfer medium may be heated by a heater attached to the heat transfer medium.
The heat transfer medium may be heated by using a composition-adjusted gas as the gas allowed to flow through a gap which is formed by the contact surface of the heat transfer medium being brought into contact with the culture vessel.
Accordingly, it possible to promote growth of cells contained in the cell suspension contained in the culture vessel.
The heat transfer medium may include a first part provided on one surface side of the culture vessel and a second part provided on the other surface side of the culture vessel opposite to the one surface. In this case, the culture vessel may be sandwiched between the first part and the second part.
Accordingly, expansion of the culture vessel due to pressurization inside the culture vessel is suppressed, and it becomes easy to control pressure inside the culture vessel. As a result, it becomes easy to control the outflow amount of the cell suspension from the culture vessel.
According to the disclosed technique, it is possible to improve a temperature rising rate of a temperature rising target and enhance a temperature uniformity of the temperature rising target.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2018-133203 | Jul 2018 | JP | national |
This application is a continuation application of International Application No. PCT/JP2019/022762 filed Jun. 7, 2019 the disclosure of which is incorporated herein by reference in its entirety. Further, this application claims priorities from Japanese Patent Application No. 2018-133203, filed Jul. 13, 2018, the disclosure of which is incorporated herein by reference in its entirety.
| Number | Date | Country | |
|---|---|---|---|
| Parent | PCT/JP2019/022762 | Jun 2019 | US |
| Child | 17123057 | US |
