JP5751856B2 - Biological cell culture container and culture apparatus - Google Patents

Description

  The present invention relates to a foldable culture vessel used when culturing living cells by a suspension method, and a culture apparatus using the foldable culture vessel.

  In the culture of living cells, it is desired to culture the living cells cheaply and safely in order to improve the productivity of the target product. However, the conventional method for culturing living cells uses a stainless steel culture tank, and a variety of incidental facilities such as a steam supply device for sterilizing the inside of the culture tank and a cleaning mechanism for cleaning are essential. Inevitably, the production cost of the target product was inevitably increased.

  Therefore, a method using single-use containers is known as a method for culturing living cells that is effective in reducing the production cost of the target product. According to this method, by using containers such as a culture tank and a medium tank for single use, a sterilization system for sterilizing them and a cleaning system for cleaning are unnecessary, and the production cost of the target product is reduced. There is expected. Further, when changing the target product, there is no fear of contamination due to the residue of the previous product, and safety can be improved.

  Patent Document 1 describes a bioreactor including a device whose wall can be deformed. Patent Document 2 describes a culture apparatus provided with a means for determining and controlling a culture state from an image of cells in a culture bag. Patent Document 3 describes a shaking device for stirring a culture solution in a culture bag. Patent Document 4 describes a disposable bioreactor in which a sensor is arranged in a liquid transfer pipe connected to the bioreactor. Patent Document 5 describes a bioreactor including a ventilation sparger, a bubble control means, a foldable bag body, and a support body that accommodates the bag body.

  Culture methods using these conventional single-use containers have been developed by adding functions essential for use as a bioreactor by applying techniques such as blood bags and infusion solution bags that have been used for many years. It has been done. While such frequent use of single-use containers greatly contributes to the reduction of system costs, on the other hand, it results in an increase in the amount of work for operators during preparations before use and removal after use. ing.

  Further, one of the problems of the culture method using these single-use containers is prevention of blockage of an exhaust filter (vent filter) provided in an exhaust pipe for exhausting exhaust gas from the culture tank. This is due to the fact that the filter membrane is covered and clogged by the mist of the condensed water or the culture solution in which the moisture contained in the exhaust gas is liquefied, and once the exhaust filter is clogged, It is difficult to continue culturing.

  Concerning this blockage of the exhaust filter, it is known that the exhaust filter can be replaced, that the filtration area of the exhaust filter is increased, and that the condensed water is prevented by heating the exhaust filter. It has been. However, these measures contribute to an increase in production cost when a larger exhaust filter is required due to an increase in culture capacity or a higher density.

  Therefore, in Patent Documents 6 to 8, the moisture contained in the exhaust gas is liquefied by applying a Peltier cooler or a water jacket to the exhaust pipe for exhausting the exhaust gas from the culture tank and cooling the exhaust gas. It is described that the exhaust filter is wetted by condensed water and culture fluid mist, that is, the moisture contained in the exhaust gas is liquefied to prevent the filter membrane from being covered and blocked by condensed water or culture fluid mist. .

JP-T-2004-535826 No. 2007-052716 No. 2007-052716 JP 2009-536520 A Special table 2009-539408 JP 2009-072182 A JP 2009-291192 A Special table 2010-525833 gazette

  However, when trying to apply a cooler consisting of a Peltier cooler or a water jacket to a foldable culture vessel for single use that has an exhaust line extending from the vessel body, the operator must be integrated with the culture vessel. The exhaust pipe must be routed and installed so as to pass through the cooler.

  In addition, a peltier cooler or a cooler comprising a water jacket is attached in advance to an exhaust pipe integral with the container body, and a single-use foldable culture container equipped with a cooler is integrated with the culture container. Although it is not necessary to arrange the exhaust pipe so as to pass through the cooling chamber of the cooler, the convenience of the foldable culture vessel is impaired, and the cooler is discarded together with the culture vessel after use. As a result, production costs increase.

  The present invention has been made in view of the above-described problems, and further improves the performance as a bioreactor by increasing the reliability in culture by preventing clogging of the exhaust filter and improving the usability. It is an object of the present invention to provide a single-use living cell culture container for single use and a culture apparatus using this culture container.

In order to solve the problems described above, the present invention is provided in a container main body formed of a foldable member, an exhaust pipe for discharging exhaust gas extending from the container main body, and the exhaust pipe. A culture apparatus for culturing living cells using a single-use culture vessel, comprising an exhaust filter that allows a flow of exhaust gas and prevents foreign substances from entering the container body from the outside, A cooling block configured to be detachable from a pipe line and a support column provided at a position where a support member of the exhaust filter is sequentially raised, and a cooling surface of the cooling block is closely attached to a wall surface of the exhaust pipe line The dew point temperature of the exhaust gas flowing into the exhaust filter is kept low as compared with the temperature of the filtration surface of the exhaust filter by cooling the wall surface of the exhaust pipe.

  According to the present invention, the exhaust filter can be prevented from being blocked, and the reliability in culture can be improved. In addition, by improving usability, it is possible to improve the efficiency of preparation work at the start of use and removal work after use. By providing a culture vessel and a culture apparatus having these effects, the culture can be continued for a long period of time, and the productivity and safety of the target product can be improved.

1 is a schematic view showing an embodiment of a biological cell culture container according to the present invention. It is the figure which showed the Example of the cooling block of the cooling device applied to the culture container which concerns on this Embodiment. It is the schematic which showed the open state before use of a cooling block. It is a schematic diagram which shows the culture apparatus to which the culture container of the biological cell which concerns on another embodiment of this invention is applied. It is a block diagram of one Example of the exhaust block unit provided with the cooling block. It is a block diagram of another Example of an exhaust block unit.

  DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of a biological cell culture container and a culture apparatus according to the present invention will be described below in detail with reference to the drawings.

  The biological cell culture vessel and culture apparatus according to the present invention can be applied to culture of cells that produce a target product as a main raw material such as pharmaceuticals. In the biological cell culture vessel and culture apparatus according to the present invention, examples of the target product to be produced include proteins such as antibodies and enzymes, physiologically active substances such as low molecular compounds, high molecular compounds, and viruses. Can do. Examples of cells to be cultured include animal cells, plant cells, insect cells, bacteria, yeasts, fungi, and algae. It is particularly suitable when animal cells producing proteins such as antibodies and enzymes are to be cultured.

FIG. 1 is a schematic view showing an embodiment of a biological cell culture vessel according to the present invention.
The culture container 10 has a container body 11 formed of a transparent foldable member such as a synthetic resin film. The container main body 11 is a usage form, for example, a cylindrical bag-like container having a diameter of 560 mm, a culture liquid filling height of 550 mm, and a culture volume of 100 L. In this usage pattern, a rigid top plate 12 made of a rigid member is fixed to a portion corresponding to the top of the container body 11 while maintaining airtightness in the container body.

  A driven portion of the stirring mechanism 20 is accommodated in the container body. The stirring mechanism 20 causes the culture solution in the container body to flow in the container body so that living cells floating in the culture solution are not deposited. The driven portion of the stirring mechanism 20 is connected to the magnetic coupling member 21 on the transmission side and extends, and the stirring blade 23 (23a, 23b) is fixed to the shaft portion 22 having rigidity. The magnetic coupling member 21 is magnetically connectable to the transmission-side magnetic coupling member 25 disposed outside the container body with the rigid top plate 12 interposed. The output shaft of the motor 26 is connected to the opposite side of the transmission side magnetic coupling member 25 opposite to the magnetic coupling member 21. The magnetic coupling member 25 constitutes a drive unit of the stirring mechanism 20 together with the motor 26. Thereby, the stirring mechanism 20 magnetically connects the magnetic coupling member 21 on the transmission side in the container body to the magnetic coupling member 25 on the transmission side outside the container body via the rigid top plate 6. In this state, if the transmission-side magnetic coupling member 25 is rotated on the rigid top plate 12 by the motor 26, the transmission-side magnetic coupling member 21 is rotated in response to the shaft portion 22 and the stirring blade 23. Is configured to rotate integrally. The airtightness of the culture vessel 10 by attaching the stirring mechanism 20 in the present embodiment is configured such that the driven part in the container main body is separated and independent from the driving part outside the container main body, and the connection and rotational force between the two are determined. It is held by using magnet couplings 21 and 25 for transmission. Thereby, stirring / mixing of the culture solution 15 stored in the container main body can be carried out with airtightness without providing the culture container 10 with a shaft through hole for transmitting the rotation of the motor 26.

  Further, a plurality of bendable piping tubes 30 (31, 32,...) Are attached to the container main body 11 while maintaining the airtightness of the attachment portions with respect to the container main body 11. The piping tube 30 constitutes an in-liquid aeration gas supply pipe 31, a gas-phase gas supply pipe 32, a liquid supply pipe 33, an exhaust circulation pipe 34, a culture liquid discharge pipe 35, and the like. In addition to this, piping tubes for sampling, injection of a drug for adjusting the pH, replacement of the medium, etc., a nozzle for taking in and out the culture solution 15 and the like are attached to the container body 11 while maintaining airtightness. The illustration is omitted for the sake of complexity.

  One end of the submerged aeration gas supply pipe 31 is led out of the main body of the culture vessel 10, and the other end is extended so as to reach the bottom side in the main body in the usage form. A connector is provided at one end of the submerged aeration gas supply pipe 31 and an output pipe of the aeration gas adjusting device 41 is connected thereto. The aeration gas regulating device 41 controls the supply of the aeration gas supplied to the culture vessel 10 and mainly controls the dissolved oxygen concentration of the culture solution 15 stored in the vessel main body to cope with the change in the dissolved oxygen concentration. Then, increase or decrease the aeration gas amount and its oxygen partial pressure. The other end of the gas supply pipe 31 for submerged aeration is connected to an air diffuser 42 arranged on the bottom side in the container body in the usage form. The air diffuser 42 discharges the aeration gas supplied from the aeration gas regulating device 41 into bubbles in the culture solution stored in the container body. The air diffuser 42 is not particularly limited, and includes a known ring sparger, or an air diffuser configured by combining a plastic porous material, a metal porous material, an inorganic porous material, or a combination thereof. Prepare. The size of the bubble to be generated is appropriately selected in advance according to the living cell to be cultured.

  One end side of the gas-phase gas supply pipe 32 is led out of the main body of the culture vessel 10, and the other end is disposed and opened on the ceiling side in the main body. A connector is provided at one end of the gas phase gas supply pipe 32, and an output pipe of the gas phase gas control device 43 is connected thereto. The gas-phase gas control device 43 controls the supply of gas-phase gas to be supplied into the container main body, and mainly controls the pH in the culture medium in the container main body, corresponding to the change in pH in the culture liquid. Increase or decrease the aeration gas amount and its carbon dioxide partial pressure.

  Gas filters 44 and 45 and on-off valves 46 and 47 are disposed in the middle of the gas supply pipe 31 for in-liquid ventilation and the gas supply pipe 32 for gas phase. The gas filters 44 and 45 prevent microorganisms from entering the container body from the outside. Moreover, each on-off valve 46 and 47 performs inflow and interruption | blocking in the container main body of the culture | cultivation container 10, and sealing the inside of a container main body, respectively of gas for aeration or gas-phase.

  One end side of the liquid supply pipe 33 is led out of the main body of the culture vessel 10, and the other end is disposed in the container main body and opened. A connector is provided at one end of the liquid supply pipe 33 so that a living cell to be cultured and its culture solution can be injected into the culture vessel 10. In the middle of the liquid supply pipe 33, an on-off valve 48 is interposed to perform inflow / blocking of the culture solution into the container body of the culture container 10 and to seal the inside of the container body.

  One end side of the exhaust circulation pipe 34 is arranged and opened on the ceiling side in the container body of the culture vessel 10, and the other end side is led out of the culture vessel 10 from the top side of the container body 11. Yes. An exhaust filter 51 is connected to the other end of the exhaust circulation pipe 34. The exhaust circulation pipe 34 is opened to the external atmosphere through the exhaust filter 51 and the gas pressure regulating valve 52 connected to the exhaust filter 51 in order, and exhausts the exhaust gas generated in the container body during the culture. The gas pressure regulating valve 52 adjusts the pressure of the gas phase in the container body, and the internal pressure of the culture container 10 is atmospheric pressure (external) in order to maintain the shape of the culture container 10 and prevent invasion of germs into the container body. Control the pressure to be slightly higher than (atmospheric pressure).

  In addition, about the gas pressure regulation valve 52, when the internal pressure of the culture container 10 is always higher than atmospheric pressure (external atmospheric pressure), the installation can be omitted. As a reason, when exhaust gas is allowed to pass through the exhaust filter 51, filtration resistance usually occurs. Therefore, if the gas is continuously injected into the vessel body of the culture vessel 10 through the submerged aeration gas supply pipe 31 and the gas phase gas supply pipe 32, an internal pressure corresponding to the filtration resistance is contained in the vessel body. Occurs. As long as the internal pressure exceeds the pressure necessary for the development and maintenance of the shape of the culture vessel 10, the gas pressure regulating valve 52 can be omitted as described above.

  One end side of the culture medium discharge pipe 35 is disposed at the bottom of the culture vessel 10 and opened at the bottom, and the other end is led out of the culture vessel 10. An opening / closing valve 49 is interposed in the middle of the culture medium discharge pipe 35 to allow the culture medium to flow out and shut off and to seal the inside of the container body.

  The culture vessel 10 having the above-described configuration has, for example, a configuration in which an exhaust filter 51 is integrally provided in an exhaust circulation pipe 34 for exhausting exhaust gas. Before use, the ends of the liquid supply pipe 33 and the culture solution discharge pipe 35 are kept sealed by, for example, plugging the connector until just before use in order to prevent invasion of microorganisms and leakage of the internal solution. In addition, the culture container 10 is in a state in which the gas in the container body is substantially removed and the container body 11 is folded. When the culture vessel 10 is folded or used, when the folded culture vessel 10 is unfolded, the container body is supported by the protruding portion of the housing member in the vessel body such as the stirring blade 23. Consideration is also given to the shape, dimensions, and material of the housing member so as not to damage 11.

  By the way, during the culture of living cells, the exhaust gas discharged from the inside of the container body of the culture container 10 contains condensed water in which moisture has been liquefied and mist of the culture solution. The moisture in the exhaust gas is saturated at about the culture temperature. That is, the dew point temperature of the exhaust gas is substantially equal to the culture temperature. Usually, culture of living cells is performed by heating the temperature of the culture solution 15 to about 25 to 50 ° C., which is higher than the environmental temperature. Therefore, during the culture of living cells, the exhaust circulation pipe 34, the exhaust filter 51, and the gas pressure regulating valve 52 that are exposed to the environment are determined by the culture temperature of the culture solution stored in the container body of the culture container 10. The surface temperature is also low. As a result, on the surfaces of the exhaust gas flow pipe 34, the exhaust filter 51, and the gas pressure regulating valve 52 that come into contact with the exhaust gas, the difference between the surface temperature and the dew point temperature of the exhaust gas from the inside of the container body is condensed. If the filter surface of the exhaust filter 51 is covered with the dew condensation water, the exhaust filter 51 is blocked, and the exhaust gas cannot be discharged from the container body. That is, it is difficult to continue the culture in the culture vessel 10.

  Therefore, in the culture vessel 10 according to the present embodiment, the cooling block 61 of the cooling device 60 is provided in a portion of the exhaust circulation pipe 34 between the vessel main body 11 exposed to the environment and the exhaust filter 51. The cooling block 61 attached to the exhaust flow pipe 34 is detachable, and the temperature of the exhaust gas discharged from the container body and flowing into the exhaust filter 51 is forcibly lowered.

  Further, with respect to the exhaust circulation pipe 34, consideration is given to the amount of heat transfer through the wall surface also in the portion where the cooling block 61 is mounted. That is, in the above-mentioned gas supply pipe 31 for aeration, gas supply pipe 32, liquid supply pipe 33, culture liquid discharge pipe 35, etc., thick silicon rubber tubes are used so as not to break during use. In contrast, a means for increasing the heat transfer coefficient using a thin-walled silicon rubber tube is provided for the portion of the exhaust flow pipe 34 where the cooling block 61 is mounted. As another tube material, a tube constructed of the same synthetic polymer material as that of the container body 11 may be used. Further, all of the exhaust circulation pipe 34 may be constructed with a tube having a large heat transfer coefficient. When a thin-walled silicon rubber tube is used, it is preferable to use a composite member whose outer peripheral surface is reinforced with a member having high mechanical strength. Further, by devising the sterilizing means, a tube made of a metal material having a larger heat transfer coefficient can be used.

FIG. 2 is a diagram showing an example of the cooling block of the cooling device applied to the culture vessel according to the present embodiment.
In the case of the present embodiment, the cooling block 61 is formed using an aluminum material having a high thermal conductivity. The cooling block 61 is a through-hole designed so that the groove surface is in contact with the outer peripheral surface of the exhaust flow pipe 34 so as to form an insertion hole 62 through which the exhaust flow pipe 34 is inserted in a mounting form with respect to the exhaust flow pipe 34. A pair of block constituent parts 61a and 61b having a groove 63 are configured to be able to be joined together with their groove forming surfaces aligned. Both block constituent portions 61a and 61b are connected to each other by a connecting member such as a hinge 64, for example, and can open and close (separate and abut) their respective joint surfaces (groove forming surfaces).

FIG. 3 is a schematic diagram showing an open state of the cooling block before use.
An inflow port 65 is formed in the lower part and an outflow port 66 is formed in the upper part on the outer peripheral surface of each of the block constituting parts 61a and 61b. A cooling channel 67 is formed in which the channel height position gradually rises toward. In addition, the inlet 65 of the block component 61a communicates with a cooling water supply port of the cooling device main body (not shown), while the outlet 66 of the block component 61a bends to the inlet 65 of the block component 61b. The flow outlet 66 of the block component 61b is connected to a cooling water recovery port (not shown) of the cooling device main body.

  As a result, the cooling water flows from the lower side to the upper side in the cooling flow path 67 formed in each of the block constituent parts 61a and 61b, and the gas mixed in the cooling water stays in the cooling flow path 67. The structure is difficult, and the groove surface of the through groove 63 of each of the block constituting portions 61a and 61b is cooled effectively and efficiently.

  Although not shown, the cooling block 61 has a configuration in which a protective cover using a heat insulating material is attached and fixed to the outer peripheral portion of the cooling block 61, that is, the outer peripheral surfaces of the block constituent portions 61a and 61b, or a detachable configuration. The cooling effect can be further improved.

  Then, when the pair of block constituting parts 61a and 61b are closed and the groove forming surfaces are brought into contact with each other, the exhaust circulation pipe 34 is inserted through the insertion hole 62 formed by the through groove 63. By holding the exhaust flow pipe 34 in the through groove 63 in advance, the cooling block 61 can be easily attached to the exhaust flow pipe 34. Further, the pair of block constituent parts 61a and 61b are opened, the groove forming surfaces are separated from each other, and the exhaust circulation pipe 34 is taken out from the through groove 63 of the pair of block constituent parts 61a and 61b. The block 61 can be easily removed from the exhaust flow pipe 34.

  In this way, by mounting the cooling block 61 to the exhaust gas flow pipe 34, the exhaust water inside the corresponding part is passed through the outer wall surface of the exhaust gas flow pipe 34 by the cooling water flowing through the cooling flow path 67. Heat can be removed locally. At that time, in the corresponding part of the exhaust gas flow pipe 34 in front of the exhaust filter 51, the temperature of the exhaust gas inside thereof decreases, so that a part of the moisture in the exhaust gas is condensed. When this moisture condenses, the moisture grows in the form of water droplets with the mist in the exhaust gas as a nucleus and adheres to the inner wall surface of the exhaust circulation pipe 34. By the operation of condensing the moisture of the exhaust gas, the mist derived from the culture solution can be removed from the exhaust gas.

  In addition, by installing the cooling block 61, the dew point temperature of the exhaust gas that is supplied to the exhaust filter 51 after removing moisture from the exhaust gas and the mist derived from the culture solution is the temperature of the exhaust gas cooled by the cooling block 61. It becomes approximately equal to temperature. As a result, the exhaust filter 51 is wetted by the mist of condensed water or culture solution in which moisture contained in the exhaust gas is liquefied, that is, the exhaust filter is filtered by mist of condensed water or culture solution in which the moisture contained in the exhaust gas is liquefied. It is possible to prevent the filter film 51 from being covered and blocked. Therefore, in the cooling device 60 having the cooling block 61 detachably attached to the exhaust circulation pipe 34, the dew point temperature of the exhaust gas flowing through the exhaust circulation pipe after being cooled by the cooling block 61 is higher than the filtration surface temperature of the exhaust filter 51. The temperature of the cooling water supplied to the cooling block 61 is set so as to be low.

  In designing the cooling block 61, it is necessary to calculate the contact area of the through groove 63 or the insertion hole 62 while paying attention to the heat transfer amount through the wall surface of the exhaust flow pipe 34.

  Further, when the diameter of the exhaust circulation pipe 34 is thin, condensed water droplets adhering to the inner wall surface may flow into the exhaust filter 51 as the exhaust gas flows. Accordingly, when determining the inner diameter of the exhaust circulation pipe 34 of the culture vessel 10, the submerged gas supply pipe 31 and the gas phase gas supply pipe 32 are provided in the container body based on the oxygen consumption rate of the living cells to be cultured. It is necessary to calculate the amount of gas injected through the gas in advance so that the gas flow rate does not become excessive in the exhaust circulation pipe 34 even at the maximum injection amount.

  Further, when the cooling block 61 is attached to the exhaust gas flow pipe 34, it is necessary to take care not to cause a liquid pool in the exhaust gas flow pipe 34. In the present embodiment, the cooling block 61, the exhaust filter 51, and the gas pressure regulating valve 52 are further provided with support members 72 (72 a, 72 b, 72 c) attached to the support column member 71 so that the height positions are sequentially increased. ) Is supported so that the flow direction of the exhaust gas by the exhaust circulation pipe 34 does not extend downward. As a result, an effect of fixing the bendable exhaust flow pipe 34 that is integrally fixed to the foldable container main body 11 while maintaining airtightness can be obtained. It is possible to prevent a liquid pool from being generated in the middle of the pipe 34 or the like.

  In the above-described embodiment, the cooling block 61 of the cooling device 60 is configured to be detachable from the exhaust circulation pipe 34 because the cost and the cooling surface do not directly contact the exhaust gas. If the cost can be solved, each culture vessel is integrally formed in advance with an exhaust circulation pipe 34 that is integrally fixed to the vessel main body 11 while maintaining airtightness. It is also possible to make it a single use as a department. In that case, the inlet 65 and the outlet 66 of the cooling block 61 may be configured such that the cooling water supply pipe and the cooling water recovery pipe extending from the cooling device main body can be attached and detached. Furthermore, although the cooling device 60 was demonstrated as a structure which distribute | circulates cooling water to the cooling block 61 and cools, the refrigerant | coolant fluid distribute | circulated to a cooling device is not restricted to cooling water. Further, the structure of the cooling block 61 is not limited to the cooling structure using the refrigerant fluid.

  According to the biological cell culture vessel 10 and the culture apparatus 100 according to the present embodiment, it is possible to prevent the exhaust filter 51 from being blocked and improve usability, thereby functioning as a single-use biological cell bioreactor for a long period of time. Can be made.

  Next, a biological cell culture vessel 10 and a culture apparatus 100 according to another embodiment of the present invention will be described with reference to the drawings.

  In the description, the same or similar components as those in the culture vessel 10 and the culture apparatus 100 according to the above-described embodiment are denoted by the same reference numerals, and detailed description thereof is omitted.

  FIG. 4 is a schematic view showing a culture apparatus to which a biological cell culture vessel according to another embodiment of the present invention is applied.

  First, in addition to the configuration described in FIG. 1, the culture container 10 according to the present embodiment has a measurement element 82 a provided with a phosphor made of a fluorescent member attached to a predetermined position on the inner wall surface of the container body 11. It is configured. In the measuring element 82a, a gap is formed between the phosphor surface of the phosphor and the inner wall surface of the affixed container body, and the culture solution 15 can be interposed. The phosphor of the measuring element 82a is excited by receiving predetermined irradiation light and emits fluorescence when returning to the ground state. In this case, the light emitted from the phosphor is affected by the measurement target in the culture solution, for example, the presence of oxygen, and the greater the amount of oxygen, the greater the attenuation of the fluorescence. In the illustrated example, the measurement element 82a constitutes an oxygen concentration measurement sensor 82 together with a detector 82b disposed opposite to the outside of the container body to which the measurement element 82a is attached. The detector 82b includes a light emitting element and a photon detection element. The light emitted from the light emitting element of the detector 82b passes through the corresponding peripheral wall portion of the container body 11 and the culture solution 15 in the predetermined gap, and irradiates the phosphor provided on the measuring element 82a. The fluorescence emitted from the measuring element 82a passes through the culture solution 15 and the peripheral wall portion of the container main body 11 in a predetermined gap, and is detected by the photon detection element of the opposing detector 82b. The detector 82b measures the fluorescence intensity received by the photon detection element and its temporal change. Therefore, the perimeter wall portion of the container body 11 to which the measurement element 82a is attached of the culture container 10 is formed of a translucent member, and excites the phosphor of the measurement element 82a from the light emitting element of the detector 82b. Therefore, the wavelength region of the fluorescence emitted from the measuring element 82a by receiving the emitted light and the reception of the emitted light is as transparent as possible.

  And the culture container 10 which concerns on this Embodiment is also in the state which the gas in a container main body was substantially extracted and folded before use. In order to prevent the culture vessel 10 from being damaged in the folded state or during the unfolding operation at the start of use, the culture vessel 10 includes the rigid top plate 12 in addition to the rigid top plate 12. Further, a support frame member 13 fixed to is provided. The support frame member 13 has a configuration in which a plate member having rigidity is bent in accordance with the outline shape of the cross section in the axial direction of the container body 11.

  In the illustrated example, the support frame member 13 has one end side that is a fixed end with respect to the rigid top plate 12, in the usage form of the container main body 11, on the tapered shoulder portion of the container main body 11 that is connected to the rigid top plate 12. Along the radial direction of the container body 11 from the rigid top 12 to the outer position of the container body 11, the other end side forming the free end is bent with respect to this one end side, along the side surface of the container body 11. In addition, the container body 11 extends toward the bottom side in parallel with the axial direction of the container body 11.

  Thus, even when a force is applied from the outside in a state where the container main body 11 is folded, it can be received by the rigid top plate 12 and the support frame member 13, and the agitation in the container main body including the container main body 11 can be performed. The driven portion of the mechanism 20, the measuring element 82a of the oxygen concentration measuring sensor 82, and the like have a structure in which the force is less likely to act directly.

  In addition, the rigid top plate 12 and the culture container 10 can be packed, transported, opened, and unfolded when the container body 11 is folded. The support frame member 13 serves to protect the culture vessel 10 by absorbing external force so that the culture vessel 10 is not damaged.

  On the other hand, the culture apparatus 100 according to the present embodiment is provided with facilities (not shown) such as gas supply equipment such as air, oxygen, nitrogen, and carbon dioxide, hot water / cold water supply equipment, and water supply / drainage equipment as necessary. Yes. For example, the gas regulating device 41 for aeration and the gas regulating device 43 for a gas phase are connected to a gas supply facility (not shown) and receive supply of component gases necessary for preparing the gas for aeration and the gas for a gas phase. For example, when the configuration of the cooling device 60 is a configuration in which cooling water is circulated through the cooling block 61 as shown in FIGS. 2 and 3, the cooling device 60 receives cooling water from a hot water / cold water supply facility (not shown). Receive supply.

  In addition, the culture apparatus 100 is provided with a measurement device for pH, temperature, dissolved oxygen, and dissolved carbon dioxide gas, and respective sensors as measurement means. In FIG. 4, only the dissolved oxygen concentration measuring device 81 and the oxygen concentration measuring sensor 82 as the dissolved oxygen measuring device are shown in the configuration, and the remaining measuring devices and sensors are not shown. For detection of pH or dissolved carbon dioxide concentration, an optical sensor similar to the configuration of the oxygen concentration measurement sensor 82 described above can be used. In that case, the phosphor used for measurement, the wavelength of the excitation light and the wavelength of the fluorescence to be measured are not particularly limited. By using a known phosphor and a combination of the wavelengths of the excitation light and the fluorescence, The objective can be achieved.

  On the other hand, the culture apparatus 100 is provided with a culture container storage case 70 that supports the culture container 10 configured as described above in a usage pattern during culture. The culture container storage case 70 is configured such that a case main body 74 that stores the culture container 10 is fixedly supported on a gantry 73. The case main body 74 has an internal shape that matches the size and shape of the container main body 11 in the usage form of the culture container 10, and in the illustrated example, the bottom part supports the bottom surface of the culture container 10 in which the culture solution 15 is stored. 74a and a bottomed cylindrical top open shape provided with a cylindrical portion 74b that supports the side surface of the culture vessel 10 so that the culture vessel 10 does not fall down. The bottom 74a has an inner bottom 74ai in which an insertion port through which the culture medium discharge pipe 35 led out from the bottom of the container main body 11 is inserted is formed in accordance with the shape of the bottom portion including the bottom of the container main body 11, and the inner bottom 74ai. And a tapered outer bottom portion 74ao connected to the bottom portion 74ai. Moreover, the cylindrical part 74b has a cylindrical shape in which the bottom side is connected to the outer edge part of the outer bottom part 74ao of the bottom part 74a. The cylindrical portion 74b is formed such that the height position of the opening end is higher than the height of the culture solution. A part of the peripheral wall of the cylindrical portion 74b becomes an openable / closable door portion 74bd, and the culture vessel 10 can be taken in and out.

  Further, the case main body 74 cooperates with an electric heater 84 for heating the culture solution 15 stored in the culture vessel 10 and a measurement element 82a disposed in the vessel main body of the culture vessel 10 to provide oxygen. A detector 82b that forms the concentration measuring sensor 82, and the like are attached.

  The electric heater 84 is fixed to the outer wall surface of the cylindrical part 74b of the case main body 74, and the generated heat can be transmitted to the culture vessel 10 through the cylindrical part 74b. Further, in order to prevent the culture vessel 10 from being damaged by the overheating, the electric heater 84 is provided with an overheat prevention control system for stopping the heating when the temperature exceeds a preset temperature.

  The detector 82b is attached to a mounting hole formed at a predetermined position of the case main body 74 via a mounting base so as to be able to be relative to the measuring element 82 arranged in the container main body of the culture vessel 10, and the light emitting element The photon detection element faces the inner peripheral surface of the cylindrical portion 74b. At this time, the holding position on the mounting base of the detector 82b can be finely adjusted. As a result, the container body 11 of the culture container 10 that is formed of a transparent foldable member such as a synthetic resin film and is in a folded form before use is slightly changed to the shape of the use form depending on the room temperature at the time of deployment. Even if the deformation of the detector 82b occurs, the light emission of the detector 82b facing the inner peripheral surface of the cylindrical portion 74b can be made by finely adjusting the holding position of the detector 82b held on the mounting base. The position and orientation of the element and the photon detection element can be moved slightly to eliminate the positional deviation that has occurred between the element and the measurement element 82a disposed in the container body 11.

  Further, the cylindrical portion 74 b of the case main body 74 has a support frame member 13 fixed to the rigid top plate 12 fixed to the top of the container main body 11 along the side surface of the container main body 11 and the container main body 11. An engaging portion 75 is formed that engages with the other end side that forms a free end extending toward the bottom side in parallel with the axial direction.

  In the illustrated example, the engaging portion 75 is formed at a predetermined position in the circumferential direction of the inner wall surface of the cylindrical portion 74b, and is parallel to the axial direction from the open end of the cylindrical portion 74b toward the bottom side of the case main body 74. The other end side of the support frame member 13 has a shape that can be fitted.

  Therefore, by engaging the other end side of the support frame member 13 fixed to the rigid top plate 12 fixed to the top part of the container body 11 with the engaging part 75 of the case body 74 in the culture container housing case 70. The culture container 10 can be accommodated by raising the support frame member 13 and the container body 11 in the case body. Further, when the container main body 11 is deployed, the measurement element 82a disposed at a predetermined position of the case main body 74 and the detector 82b disposed on the case main body 74 of the culture container housing case 70 have their fluorescence. The culture vessel 10 can be positioned and accommodated with respect to the culture vessel accommodation case 70 so that the body, the light emitting element, and the photon detection element face each other. In this case, in the case main body 74, the arrangement positional relationship between the engaging portion 75 in the circumferential direction of the inner wall surface of the cylindrical portion 74b and the detector 82b of the oxygen concentration measurement sensor 82 is the culture container in the developed usage form. 10, the arrangement positional relationship in the circumferential direction of the outer wall surface of the container body 11 between the other end side of the support frame member 13 and the measurement element 82a of the concentration measurement sensor 82 is applied.

  Therefore, the support frame member 13 is accommodated in the culture container by engaging with the engaging portion 75 of the case main body 74 as a positioning member in an operating state where the culture container 10 is supported by the culture container housing case 70 in the usage state. It functions as a part of the case 70. Further, a support column member 71 for supporting the exhaust flow pipe 34 in the culture vessel 10 can be integrally attached and fixed to the case main body 74 or the gantry 73 of the culture vessel storage case 70.

  In addition, the detector 82 b of the oxygen concentration measurement sensor 82 is connected to the dissolved oxygen concentration measurement device 81. The dissolved oxygen concentration measuring device 81 calculates the dissolved oxygen concentration of the culture solution 15 stored in the container body based on the measured value supplied from the detector 82b, and uses the calculated value as the control device of the culturing device 100. To 110.

  The control device 110 includes a measurement value from the dissolved oxygen concentration measurement device 81 and other measurement outputs such as pH and dissolved carbon dioxide concentration, which are not shown in the drawing, in advance according to the culture output and the like. Various set values such as culture time set by the input device are input. Based on these inputs, the control device 110 controls the operation of each part of the device such as the motor 26, the gas adjusting device 41 for ventilation, the gas adjusting device 43 for the gas phase, the gas pressure regulating valve 52, the cooling device 60, the electric heater 84, and the like. .

  In the biological cell culture operation using the biological cell culture container 10 and the culture apparatus 100 according to the present embodiment, the user expands the culture container 10 in a state in which the container body 11 is folded into a use form. First, the door 74bd is opened, the culture container 10 is accommodated in the culture container accommodation case 70, and the support frame member 13 of the culture container 10 is engaged with the engagement portion 75 of the culture container accommodation case 70 in the culture apparatus 100. Thus, the culture vessel 10 is positioned and held with respect to the culture vessel storage case 70 provided with the detector 82b.

  The submerged aeration gas supply pipe 31, the gas phase gas supply pipe 32 and the like are connected in communication with the aeration gas adjustment apparatus 41, the gas phase gas adjustment apparatus 43 and the like, and the culture medium discharge pipe 35 is connected to the culture vessel. Derived from the storage case 70. A cooling block 61 is attached to the exhaust flow pipe 34, and the exhaust filter 51 is connected to the gas pressure regulating valve 52 as necessary. The cooling block 61, the exhaust filter 51, and the gas pressure regulating valve 52 are supported by the support column member 71 by the support members 72a to 72c, and the flow direction of the exhaust gas flow path by the exhaust flow pipe 34 does not extend downward. In this way, the exhaust flow pipe 34 is prevented from bending so as to cause a liquid pool. At this time, the plurality of bendable tube tubes 30 of the culture vessel 10 including the gas supply pipe 31 for gas passage in the liquid, the gas supply pipe 32 for the gas phase, and the like prevent invasion of microorganisms and leakage of the internal solution. Therefore, the sealed state is maintained until just before use.

  The user magnetically couples the magnetic coupling member 21 in the container body of the culture vessel 10 to the magnetic coupling member 25 with the rigid top 12 interposed therebetween, and rotates the motor 26 to the magnetic coupling member 25. Connect shafts. In this case, the motor 26 is fixedly supported on the gantry 73 via a mounting base (not shown) whose mounting position can be freely changed, so that vibration associated with its weight and rotational drive does not directly act on the culture vessel 10. Yes. Thereby, the culture vessel 10 is released from the weight of the motor 26 and the drive vibration.

  The user closes the door 74bd opened for accommodating the culture vessel 10 and holds and fixes the culture vessel 10 in the culture vessel accommodating case 70. Thereafter, the gas was introduced into the culture vessel 10 from the aeration gas regulating device 41 or the gas phase gas regulating device 43 via the submerged aeration gas supply pipe 31 or the gas phase gas supply pipe 32, thereby being folded. The culture container 10 which is in a state can be developed and used.

  At this time, the deployed culture container 10 is connected to the peripheral wall of the container body 11 to which the measuring element 82a is attached by the engagement between the support frame member 13 of the culture container 10 and the engaging portion 75 of the culture container storage case 70. The portion faces the portion where the detector 82b of the culture container housing case 70 is disposed and is brought into close contact therewith. Thus, during culture, the fluorescent screen of the measuring element 82a and the photon detection element of the detector 82b are containers other than the excitation light from the light emitting element of the detector 82b and the fluorescence from the fluorescent plane of the measuring element 82a. A structure in which external light from the outside of the main body is difficult to enter can be obtained.

  The user uses the liquid supply pipe 33 to inject the living cells and the culture solution into the culture vessel 10 when culturing the living cells. At that time, when living cells to be cultured have anchorage dependency, microcarriers serving as a scaffold for proliferation are mixed and used in the culture solution. Thereby, the living body cell which has anchorage dependence can adhere to a culture surface, and can be propagated by contacting a microcarrier.

  Then, when the culture of the living cells is started by the user's start operation, the aeration gas regulating device 41 and the gas phase gas regulating device 43 are controlled by the control device 110 during the cultivation. And the amount of aeration gas are controlled, and the dissolved oxygen concentration in the gas phase of the culture vessel 10 and the pH of the culture solution are controlled. The gas pressure regulating valve 52, the motor 26, the electric heater 84, and the cooling device 60 are also controlled by the control device 110 to control the internal pressure of the culture vessel 10, the stirring speed of the culture solution, the heating of the culture solution, and the cooling of the exhaust gas. Control is done.

  The control of each part described above is based on the output of various detectors such as the oxygen concentration measurement sensor 82, pH, dissolved carbon dioxide concentration, etc. (not shown), and various preset values set in advance according to the culture target, etc. This is performed according to a procedure registered in advance in the storage unit of the control device 110.

  Here, the control of the dissolved oxygen concentration in the culture solution will be described as an example. A method of increasing or decreasing the stirring rotation speed of the stirring blade 23 in the stirring mechanism 20 by controlling the motor 26, the gas adjusting device 41 for aeration, and the gas phase A method of controlling the gas regulating device 43 to increase or decrease the amount of oxygen-containing gas blown into the submerged gas supply pipe 31 or gas-phase gas supply pipe 32 or the oxygen partial pressure, or a method of using these methods in combination. The control device 110 performs control based on the measurement value supplied from the dissolved oxygen concentration measurement device 81. For example, in the method in which the control device 110 increases or decreases the stirring rotation speed of the stirring blade 23 in the stirring mechanism 20, when the stirring rotation speed of the stirring blade 23 in the stirring mechanism 20 increases, the stirring / mixing of the culture solution 15 is strengthened and the culture is performed. As the amount of dissolved oxygen in the liquid 15 increases, the floating state of living cells in the culture solution is improved.

  In addition, control by the cooling block 61 of the cooling device 60 to lower the dew point temperature of the exhaust gas flowing in the exhaust flow pipe to be lower than the filtration surface temperature of the exhaust filter 51 is performed by the control device 110 that is detected and input. For example, the control is performed by instructing the cooling device 60 to control the temperature, the flow rate, or the like of the cooling water flowing through the cooling block 61 based on the temperature or the like.

  As described above, according to the biological cell culture vessel 10 and the culture apparatus 100 according to the present embodiment, the exhaust filter 51 can be prevented from being blocked by dew condensation of the exhaust gas, and can function as a bioreactor for a long time. A highly reliable living cell culture vessel and culture apparatus for single use can be provided.

  Next, a modified example of the cooling block 61 of the cooling device 60 configured to be detachable from the exhaust circulation pipe 34 applied to the above-described biological cell culture vessel 10 and culture device 100 will be described with reference to the drawings. explain.

  In the description, the same or similar configurations as those of the cooling block 61 shown in FIGS. 2 and 3 are denoted by the same reference numerals, and detailed description thereof is omitted.

FIG. 5 is a configuration diagram of an embodiment of an exhaust block unit including a cooling block.
In the present embodiment, the exhaust block unit 120 has a configuration in which the same cooling block 61 shown in FIGS. 2 and 3 is integrally fixed to a unit protection case 121 formed of a heat insulating material (heat insulating material). It has become. Further, in the case of the illustrated exhaust block unit 120, in addition to the exhaust flow pipe 34, the exhaust filter 51 and the flow pipe 53 on the outlet side thereof can be integrally accommodated and held. Therefore, the unit protection case 121 is attached to the exhaust gas flow pipe 34, and the exhaust gas flow pipe 34 and the outlet side flow pipe 53 are inserted to accommodate and hold the exhaust filter 51.

  The unit protective case 121 has a pair of case constituent parts 121a and 121b corresponding to the pair of block constituent parts 61a and 61b of the cooling block 61, and an exhaust gas flow pipe is provided at the joint surface of each case constituent part 121a and 121b. Stepped through holes in which insertion parts 122a1 and 122b1, block component fixing parts 122a2 and 122b2, exhaust flow pipe insertion parts 122a3 and 122b3, exhaust filter housing parts 122a4 and 122b4, and outlet side flow pipe insertion parts 122a5 and 122b5 are connected in series. Grooves 122a and 122b are formed. In addition, block constituent parts 61a and 61b of the cooling block 61 are fixed in advance to the block constituent part fixing parts 122a2 and 122b2. Both case components 121a and 121b are connected to each other by a connecting member such as a hinge 123, for example, and the respective stepped through grooves 122a and 122b are combined to open and close (ie, separate and abut) the respective joint surfaces. It can be done.

  Further, the case components 121a and 121b are formed with an inlet 124 and an outlet 125 that communicate with the inlet 65 and the outlet 66 shown in FIG. 3 of the block components 61a and 61b, respectively. The inlet 124 of the case component 121a communicates with a cooling water supply port of a cooling device main body (not shown), while the outlet 125 of the case component 121a is a tube having flexibility with respect to the inlet 124 of the case component 121b. 126, the outlet 125 of the case component 121b is connected to a cooling water recovery port of a cooling device main body (not shown). As a result, the cooling water flows through the cooling flow path 67 (see FIG. 3, not shown in FIG. 5) of the block constituent parts 61a and 61b of the cooling block 61, and the through grooves 63 of the respective block constituent parts 61a and 61b. The groove surface (cooling surface) is cooled. At that time, the housing surfaces other than the joint surfaces of the block components 61a and 61b are protected by the unit protective case 121 formed of a heat insulating material, so that the groove surface (cooling surface) of the through groove 63 is effective and efficient. The cooling effect of the cooling block 61 can be further improved.

  In addition, the exhaust filter housing portions 122a4 and 122b4 in which the exhaust filter 51 is housed and held are provided with electric heaters 127 (127a and 127b), so that the temperature of the filter surface of the exhaust filter 51 can be reduced even if the environmental temperature decreases. Is controlled by the control device 110 so as not to drop below the dew point temperature of the exhaust gas. At that time, it is preferable that the atmosphere temperature of the housing space of the exhaust filter 51 formed by the exhaust filter housing portions 122a4 and 122b4 is equal to or higher than the temperature of the culture solution. The electric heater 127 is provided with an independent overheat prevention mechanism (not shown) so as not to exceed the allowable temperature of the exhaust filter 51.

  According to the present embodiment, since the cooling block 61 and the exhaust filter 51 are detachably attached to the exhaust block unit 120 and are collectively accommodated and fixed, the usability is improved. Moreover, since the temperature of the exhaust filter 51 can be maintained at a temperature equal to or higher than the temperature of the culture solution 15, the risk of causing the exhaust filter 51 to be blocked can be further reduced, and the culture can be maintained for a long period of time and the product safety can be improved.

FIG. 6 is a configuration diagram of another embodiment of an exhaust block unit including a cooling block.
The exhaust block unit 120 of the present embodiment is configured to directly cool the cooling block 61 using the Peltier element 128 using the Peltier effect without depending on the flow of the cooling water, so that the space in which the exhaust filter 51 is accommodated. The heat from the heat generating surface of the Peltier element 128 can also be used for heating. In the description, the same or similar components as those in the exhaust block unit 120 shown in FIG. 5 are denoted by the same reference numerals, and detailed description thereof is omitted.

  Also in the present embodiment, the exhaust block unit 120 has a configuration in which the cooling block 61 is integrally disposed in a unit protection case 121 formed of a heat insulating material (heat insulating material). Note that the cooling block 61 of the present embodiment does not need to include the inlet 65, the outlet 66, and the cooling channel 67 related to the circulation of the cooling water, and is formed using a member having high thermal conductivity. Just do it.

  Therefore, the unit protective case 121 has a pair of case constituent parts 121a and 121b corresponding to the pair of block constituent parts 61a and 61b of the cooling block 61, and the joint surfaces of the case constituent parts 121a and 121b are provided with exhaust gas. Stepped through grooves 122a and 122b are formed in which the flow pipe insertion portions 122a1 and 122b1, the cooling block / exhaust filter housing portions 122a6 and 122b6, and the outlet side flow pipe insertion portions 122a5 and 122b5 are connected.

  The cooling block / exhaust filter housing portions 122a6 and 122b6 of the pair of block constituting portions 61a and 61b are fixedly provided with the block constituting portions 61a and 61b of the cooling block 61 on the exhaust circulation pipe insertion portions 122a1 and 122b1, respectively. The exhaust filter 51 is accommodated on the side circulation pipe insertion portions 122a5 and 122b5 side. In the illustrated example, the Peltier element 128 has a heat generating surface directed to the filter housing space 130 formed by the cooling block / exhaust filter housing portions 122a6 and 122b6, and the cooling surface of the block constituting portions 61a and 61b of the cooling block 61. Are fixed to the block constituent parts 61a and 61b in contact with the filter side end face. Further, a heat radiating member 129 for heat dissipation is affixed to the heat generating surface of the Peltier element 128 so that heat can be efficiently radiated to the atmosphere of the filter housing space 130. The operation of the Peltier element 128 is controlled by the control device 110 via a drive circuit (not shown).

  In the present embodiment, an exhaust fan 131 and a ventilation passage 132 for exchanging the atmosphere of the filter housing space 130 with the atmosphere of the external environment are communicated with the filter housing space 130 in the case component 121a of the unit protection case 121. Is provided. The operation of the exhaust fan 131 is also controlled by the control device 110.

  In the exhaust block unit 120 of the present embodiment, the Peltier element 128 is operated in a mounting form with respect to the exhaust circulation pipe 34 in which the joint surfaces of the pair of case components 121a and 121b of the unit protective case 121 are joined. Thus, the pair of block components 61a and 61b of the cooling block 61 can be cooled by the Peltier effect, and the dew point temperature of the exhaust gas flowing through the exhaust circulation pipe can be made lower than the filtration surface temperature of the exhaust filter 51. . At the same time, the heat generated from the heat generating surface of the Peltier element 128 at that time is dissipated from the heat radiating member 129 toward the filter housing space 130, and therefore the temperature of the filter surface of the exhaust filter 51 may be lower than the dew point temperature of the exhaust gas. Can be warmed so that there is no. At that time, if the atmosphere of the filter housing space 130 is overheated, the exhaust fan 131 is operated to cool the air by changing the atmosphere, the temperature of the filter housing space 130 is adjusted, and the filter surface of the exhaust filter 51 is overheated. Can be prevented.

  As shown in the figure, when the cooling capacity of the cooling block 61 is insufficient only by arranging the Peltier elements 128 on the filter side end faces of the block constituent parts 61a and 61b, the joint surfaces of the block constituent parts 61a and 61b. It is also possible to arrange the Peltier element 128 with its cooling surface in contact with other surfaces. In this case, since the heat generating surfaces of these Peltier elements 128 cannot directly face the filter housing space 130, for example, the case components 121a and 121b are provided with air ducts communicating with the filter housing space 130 to operate the exhaust fan 131. Thus, the heat generation surface can be guided to the filter housing space 130 to heat the exhaust filter 51.

  According to the present embodiment, since the cooling block 61 and the exhaust filter 51 are detachably attached to the exhaust block unit 120 and are collectively accommodated and fixed, the usability is improved. Furthermore, the cooling water piping for cooling the cooling block 61 is not required, and the usability is improved.

10 culture vessel, 11 vessel body, 12 rigid top plate, 13 support frame member,
15 culture solution, 20 stirring mechanism, 21 magnetic coupling member, 22 shaft,
23 stirring blade, 25 magnetic coupling member, 26 motor,
30 piping tube, 31 gas supply pipe for venting in liquid, 32 gas supply pipe for gas phase,
33 Liquid supply pipe, 34 Exhaust flow pipe, 35 Culture medium discharge pipe,
41 gas regulator for ventilation, 42 air diffuser, 43 gas regulator for gas phase,
44, 45 Gas filter, 46, 47, 48, 49 On-off valve,
51 exhaust filter, 52 gas pressure regulating valve, 60 cooling device,
61 cooling block, 62 insertion hole, 63 through groove, 64 hinge,
65 inlet, 66 outlet, 67 cooling flow path, 68 tube,
70 culture vessel storage case, 71 support column member, 72 support member, 73 mount,
74 case main body, 75 engaging part, 81 dissolved oxygen concentration measuring device,
82 oxygen concentration measuring sensor, 82a measuring element, 82b detector,
84 Electric heater, 100 culture device, 110 control device,
120 exhaust block unit, 121 unit protective case,
122 stepped through groove, 123 hinge, 124 inlet, 125 outlet,
126 tubes, 127 electric heaters, 128 Peltier elements,
129 heat radiating member, 130 filter housing space, 131 exhaust fan,
132 Ventilation passage.

Claims (8)

A container body formed of a foldable member;
An exhaust pipe for discharging exhaust gas extending from the container body;
An exhaust filter that is provided in the exhaust pipe line, allows the exhaust gas to flow, and prevents foreign matter from entering the container main body from the outside;
A culture apparatus for culturing living cells using a single-use culture vessel equipped with
A cooling block configured to be detachably attached to the exhaust pipe, and a support column provided at a position where the support member of the exhaust filter is sequentially raised,
The cooling surface of the cooling block is brought into close contact with the wall surface of the exhaust pipe line to cool the wall surface of the exhaust pipe line, whereby the exhaust gas flowing into the exhaust filter is compared with the temperature of the filtration surface of the exhaust filter. A culture apparatus characterized by maintaining a low dew point temperature. The culture apparatus according to claim 1, wherein the cooling block has a unit structure in which the exhaust pipe line and the exhaust filter are provided at sequentially increasing positions and are integrally accommodated and held. The cooling block includes a Peltier element.
The cooling block surrounds the outer wall surface of the exhaust pipe line with the cooling surface located on the heat absorption side of the Peltier element in the closed state, and releases the enclosure by the cooling surface in the open state to release the exhaust pipe line The culture apparatus according to claim 1, further comprising an attaching / detaching mechanism for releasing the sucrose. The cooling fluid is circulated through the flow path formed in the cooling block,
The cooling block includes an attachment / detachment mechanism that surrounds an outer wall surface of the exhaust pipe line with the cooling surface in the closed state, and releases the exhaust pipe line by releasing the enclosure by the cooling surface in the open state. The culture apparatus according to claim 1 or 2, wherein 4. A heating device is provided that keeps the temperature of the filtration surface of the exhaust filter higher than the culture temperature of living cells by air heated by heat dissipation on the heat generation side of the Peltier element. The culture apparatus described in 1.   The heating device includes an air flow rate adjusting unit that adjusts the flow rate of air supplied to the heat generating side of the Peltier element in order to adjust the temperature of the filtration surface of the exhaust filter. The culture apparatus according to claim 5.   The culture apparatus according to claim 1 or 4, further comprising a heating device that maintains a temperature of a filtration surface of the exhaust filter higher than a culture temperature of living cells.   The culture apparatus according to claim 7, wherein the heating apparatus is an electric heater type heating apparatus.

Images