WO2018047679A1

WO2018047679A1 - Culture medium heating device and culturing system

Info

Publication number: WO2018047679A1
Application number: PCT/JP2017/030981
Authority: WO
Inventors: 聡文北原; 邦忠畑林; 久五味
Original assignee: 東京エレクトロン株式会社
Priority date: 2016-09-09
Filing date: 2017-08-29
Publication date: 2018-03-15

Abstract

The culture medium heating device according to the present invention comprises: a cylindrical device body having a central axis along the vertical direction; a device inlet which is provided in one of an upper part and a lower part of the device body, and into which a culture medium flows; a device outlet which is provided in the other of the upper part and the lower part of the device body, and out of which the culture medium flows; and a device channel through which the culture medium passes from the device inlet toward the device outlet. Culture medium passing through the device channel is heated by a heating unit. The device channel is formed in a spiral shape centered on the central axis of the device body.

Description

Medium heating device and culture system

The present invention relates to a medium heating apparatus and a culture system.

In recent years, research and development of regenerative medicine that artificially creates target tissues and organs by cell culture has been promoted. In order to perform a cell culture operation, a culture system satisfying a predetermined standard, for example, GMP (Good Manufacturing Practice) is used.

In a culture system, in order to prevent the culture environment in the culture vessel from gradually deteriorating, the liquid medium (also referred to as culture solution) in the culture vessel is regularly replaced. At this time, by supplying a new medium to the culture container, the old medium in the culture container is pushed out from the culture container and discharged, and the medium in the culture container is replaced.

By the way, the new medium before being supplied to the culture container is stored at a low temperature of about 4 ° C. in a cool box or the like in order to prevent deterioration of the components. On the other hand, the medium for cell culture in the culture vessel is maintained at a high temperature of about 37 ° C. Therefore, a culture apparatus configured to heat a new medium before supplying it to the culture vessel is known (see, for example, Patent Documents 1 and 2). In such a culture apparatus, a new medium is temporarily stored in a tank or a medium supply pipe (hereinafter referred to as a tank or the like) and heated by a heater. And the culture medium which became high temperature is supplied to a culture container in the case of medium replacement | exchange.

Japanese Patent Laid-Open No. 2015-109877 Japanese Patent Laid-Open No. 2015-213462

However, it takes a considerable amount of time to heat the medium supplied at about 4 ° C. to about 37 ° C. Since the components of the medium at a high temperature are likely to deteriorate, it is preferable that the time for heating the medium is short.

On the other hand, if the medium remains in the tank or the like after the medium is discharged, the component concentration of the medium to be supplied next may fluctuate. In addition, even when the cleaning liquid remains in the tank or the like after cleaning with a cleaning liquid (for example, pure water), the component concentration of a new culture medium supplied thereafter may vary.

The present invention has been made in consideration of such points, and provides a medium heating apparatus and a culture system that can shorten the time for heating the medium and prevent the liquid from remaining.

The present invention is a culture medium heating apparatus for heating a culture medium for cell culture supplied from a culture medium supply source to a culture vessel, a cylindrical apparatus body having a central axis along the vertical direction, an upper part of the apparatus body, A device inlet provided in one of the lower portions, into which the culture medium flows, a device outlet provided in the other of the upper and lower portions of the device main body, through which the culture medium flows out, and from the device inlet. An apparatus passage through which the culture medium toward the apparatus outlet passes, and a heating unit that heats the culture medium that passes through the apparatus passage, and the apparatus passage is formed in a spiral shape around the central axis of the apparatus main body. A formed medium heating device is provided.

In the above-described culture medium heating apparatus, the apparatus inlet may be provided in an upper part of the apparatus main body, and the apparatus outlet may be provided in a lower part of the apparatus main body.

In the culture medium heating apparatus described above, a spiral groove is provided on an outer peripheral portion of the apparatus main body, the spiral groove is formed in a spiral shape around the central axis of the apparatus main body, and the apparatus passage includes the spiral groove. It may be constituted by a tube member inserted in

In the culture medium heating apparatus described above, the tube member may be pressed from the wall surface of the spiral groove.

The medium heating apparatus described above may further include a pressing member that presses the tube member from the outer peripheral side.

In the culture medium heating apparatus described above, the heating unit extends from the apparatus inlet to the apparatus outlet along the central axis of the apparatus body, and is concentrically disposed in the apparatus passage. You may do it.

The above-described culture medium heating apparatus may further include a temperature measurement unit that measures the temperature of the apparatus main body.

In the culture medium heating apparatus described above, the temperature measurement unit may be arranged in a part of the apparatus main body on the apparatus outlet side.

In the culture medium heating device described above, the temperature measurement unit may be arranged between the device passage and the heating unit.

In addition, the present invention is provided between a culture medium supply source for supplying a new culture medium, a container holding unit capable of holding a culture container, and the culture medium supply source and the container holding unit. There is provided a culture system comprising the above-described culture medium heating device for heating the culture medium supplied to a container.

The culture system described above may further include a buffer tank that is provided between the medium heating device and the container holding unit and stores the medium heated by the medium heating device.

The culture system described above may further include a culture medium supply pump that is provided between the culture medium supply source and the culture medium heating apparatus and supplies the culture medium from the culture medium supply source to the culture medium heating apparatus.

In the culture system described above, the culture system further includes a control unit that controls the heating unit of the culture medium supply pump and the culture medium heating device, and the culture medium heating device includes a temperature measurement unit that measures the temperature of the device main body, The controller may control the medium supply pump and the heating unit based on the temperature of the apparatus main body measured by the temperature measurement unit while driving the medium supply pump.

The culture system described above may further include a cleaning liquid supply source that supplies a cleaning liquid to the apparatus inlet or the apparatus outlet provided in the upper part of the apparatus main body of the medium heating apparatus.

In the culture system described above, the apparatus further includes a gas supply source that supplies a gas capable of purging the cleaning liquid to the apparatus inlet or the apparatus outlet provided in the upper part of the apparatus main body of the medium heating apparatus. May be.

According to the present invention, the time for heating the culture medium can be shortened and the liquid can be prevented from remaining.

FIG. 1 is a diagram showing a schematic configuration of a culture system in the present embodiment. FIG. 2 is a plan view showing the culture vessel shown in FIG. FIG. 3 is a cross-sectional view showing the culture container shown in FIG. FIG. 4 is a front view showing the arrangement of the inlet heater of FIG. FIG. 5 is a cross-sectional view showing the inlet heater of FIG. 6 is a partial cross-sectional view showing the inlet heater of FIG.

Hereinafter, an embodiment of the present invention will be described with reference to the drawings. Note that, in the drawings attached to the present specification, for the sake of easy understanding of the drawings, the scale and the vertical / horizontal dimension ratio are appropriately changed and exaggerated from those of the actual ones.

The culture system according to the present embodiment can be used for culturing all types of cells, including pluripotent stem cells such as (human) iPS cells and (human) ES cells, and chondrocytes such as bone marrow stromal cells (MSC). It can be used when culturing various cells such as dendritic cells. In the present embodiment, the following description will be given mainly assuming the use of culturing iPS cells, but this is only an example.

First, a schematic configuration of a culture system according to an embodiment of the present invention will be described with reference to FIG.

As shown in FIG. 1, the culture system 1 is held in a medium supply source 2 that supplies a new medium, a container holding unit 3 that can hold a culture container 100 for culturing cells, and a container holding unit 3. A medium analysis unit 4 that analyzes the components of the medium discharged from the culture vessel 100.

The medium supply source 2 stores a new medium for cell culture to be supplied to the culture vessel 100. The culture medium supply source 2 is provided, for example, in a cold storage box, and during storage, the culture medium is stored at a low temperature (for example, about 4 ° C.) to prevent deterioration of components.

Between the culture medium supply source 2 and the container holding unit 3, an inlet heater 5 (medium heating apparatus) and a new solution buffer tank 6 are provided in this order.

The inlet heater 5 heats the culture medium supplied from the culture medium supply source 2 to the culture vessel 100 to raise the temperature of the culture medium (for example, about 37 ° C.). The heated medium is discharged from the inlet heater 5 and supplied to the new solution buffer tank 6. Details of the inlet heater 5 will be described later. An inlet pump 7 (medium supply pump) is provided between the culture medium supply source 2 and the inlet heater 5 to supply a culture medium from the culture medium supply source 2 to the new liquid buffer tank 6 via the inlet heater 5. It has been. The pump for supplying the culture medium (medium supply pump) is not limited to the inlet pump 7. For example, a common pump (not shown) is provided downstream from the junction of the line from the culture medium supply source 2 and the line from the cleaning liquid supply source 13 (described later) and upstream from the inlet heater 5. The common pump may have a medium supply function. In this case, the shared pump can also have a cleaning liquid supply function, and a cleaning pump 14 described later can be omitted.

The new solution buffer tank 6 stores the medium heated by the inlet heater 5 and removes bubbles in the medium. The new solution buffer tank 6 has an internal space for storing the culture medium and a vent portion (both not shown). The internal space of the new solution buffer tank 6 is the same as that of the new solution buffer tank 6. It communicates with the surrounding atmosphere (clean atmosphere in the chamber of the culture system 1) via the vent portion. As a result, when bubbles are mixed in the medium stored in the buffer tank 6 for fresh solution, the bubbles are lifted and removed from the medium. That is, the dissolved gas and minute bubbles contained in the new medium are expressed and expanded when the temperature becomes high, and thus the bubbles can be efficiently removed from the medium stored in the internal space. In addition, since the vent portion is provided, it is possible to smoothly supply and discharge the culture medium to and from the fresh solution buffer tank 6. In addition, a vent filter (not shown) is provided in the vent portion to prevent foreign matter from entering the internal space of the new liquid buffer tank 6.

The storage capacity of the culture medium in the internal space of the new solution buffer tank 6 is such that when the culture medium in the culture container 100 is replaced, the old culture medium in the culture container 100 can be pushed out and completely replaced with a new culture medium. It is preferable to make it larger than the capacity. For example, when the capacity of the culture vessel 100 is 18 mL, the medium storage capacity of the new solution buffer tank 6 may be 30 mL, which is larger than this.

Between the inlet pump 7 and the inlet heater 5, a first inlet on-off valve 8 is provided. The first inlet open / close valve 8 controls the supply of the culture medium from the culture medium supply source 2 to the fresh solution buffer tank 6. A second inlet on-off valve 9 is provided between the new liquid buffer tank 6 and the container holding unit 3. The second inlet open / close valve 9 controls the supply of the medium from the fresh solution buffer tank 6 to the culture vessel 100. A solution discharge line (not shown) may be provided downstream of the second inlet opening / closing valve 9. The new solution buffer tank 6 is disposed at a higher position than the container holding unit 3, and the medium can be easily supplied from the new solution buffer tank 6 to the culture vessel 100 held in the container holding unit 3. It has become.

The container holding unit 3 is configured to hold the culture container 100. When the culture system 1 incorporates an incubator for cell culture (not shown), the incubator is disposed in the chamber of the culture system 1 described above, and the container holding unit 3 is provided in the casing of the incubator. It is done. This housing is configured to adjust at least one of the temperature, humidity, and gas concentration of the internal atmosphere. For example, the temperature of the atmosphere in the housing is adjusted so that the temperature of the culture vessel 100 held in the vessel holding unit 3 is about 37 ° C. On the other hand, when the culture system 1 and the incubator are installed apart from each other, the container holding unit 3 temporarily holds the culture container 100 transported from the incubator to the container holding unit 3 in order to exchange the medium. For that. In this case, the culture container 100 held by the container holding unit 3 is accommodated in a housing that maintains a sterilization space in the chamber of the culture system 1, for example. Details of the culture vessel 100 will be described later.

Between the container holding unit 3 and the medium analysis unit 4, an outlet pump 10, a medium filter 11, and an outlet opening / closing valve 12 are provided in this order. Among these, the outlet pump 10 pulls out the culture medium after cell culture from the culture vessel 100 and supplies the culture medium to the culture medium analysis unit 4. At this time, the supply of a new medium from the fresh solution buffer tank 6 to the culture container 100 is prompted simultaneously with the withdrawal of the medium from the culture container 100. As a result, a new medium is supplied to the culture vessel 100 and the medium is exchanged. The culture medium filter 11 is configured to remove solids (for example, cultured cells) contained in the culture medium discharged from the culture container 100 from the culture medium. The outlet on-off valve 12 is configured to control the supply of the medium from the culture vessel 100 to the medium analysis unit 4.

The culture system 1 according to the present embodiment further includes a cleaning liquid supply source 13. The cleaning liquid supply source 13 stores a cleaning liquid (for example, pure water) to be supplied to a heater inlet 31 (described later) of the inlet heater 5. The cleaning liquid supplied to the heater inlet 31 passes through a heater passage 33 (described later) of the inlet heater 5 and is supplied to the new liquid buffer tank 6 so that the inlet heater 5 and the new liquid buffer tank 6 are cleaned. . A cleaning pump 14 and a cleaning on / off valve 15 are provided in this order between the cleaning liquid supply source 13 and the inlet heater 5. Among these, the cleaning pump 14 is configured to supply the cleaning liquid from the cleaning liquid supply source 13 to the inlet heater 5 and the buffer tank 6 for new liquid. The cleaning on / off valve 15 controls the supply of cleaning liquid from the cleaning liquid supply source 13 to the inlet heater 5.

The culture system 1 further includes a gas supply source 16. The gas supply source 16 supplies compressed gas (for example, compressed clean air) to the heater inlet 31 of the inlet heater 5. The compressed gas supplied to the heater inlet 31 passes through the heater passage 33 of the inlet heater 5 and is supplied to the new liquid buffer tank 6, and the cleaning liquid that can remain in the inlet heater 5 is purged. A gas on-off valve 17 is provided between the gas supply source 16 and the inlet heater 5 to control the supply of compressed gas from the gas supply source 16 to the inlet heater 5. Note that the gas supply source 16 stores compressed gas, and the gas supply source 16 has a configuration in which the compressed gas is supplied to the inlet heater 5 by the pressure. The cleaning gas is purged into the inlet heater 5. As long as possible gas can be supplied, the configuration is not limited to this.

As shown in FIG. 1, the culture system 1 further includes a control unit 18. The control unit 18 is configured to control each on-off valve, pump, and heater body 37 (described later) of the inlet heater 5 described above.

Next, the culture vessel 100 according to the present embodiment will be described with reference to FIGS.

As shown in FIGS. 2 and 3, the culture vessel 100 includes a vessel body 101 and a flat plate 102 attached to one surface of the vessel body 101. The container body 101 includes a container inlet 103 into which a medium (a suspension in which cells are dispersed in addition to the medium, a release agent, phosphate buffered saline (PBS), etc.) is flowed, and a container inlet 103 A container passage 104 through which the culture medium passes, and a container outlet 105 through which the culture medium that has passed through the container passage 104 flows out. Among these, the container inlet 103 is connected to the above-described new solution buffer tank 6, and the container outlet 105 is connected to the culture medium analyzer 4.

The container passage 104 of the container body 101 is formed in a groove shape on the one surface side where the flat plate 102 of the container body 101 is attached. The diameter of the container passage 104 (that is, the depth and width of the groove) is, for example, 2 mm to 4 mm. Further, the container passage 104 of the container body 101 has a portion that meanders in plan view, that is, a portion in which straight portions and folded portions are alternately connected. Thereby, the full length of the container channel | path 104 is extended without enlarging the container main body 101, and the elongate container channel | path 104 is formed.

As shown in FIGS. 2 and 3, a plurality of cell seeding regions 106 in which cells passing through the container passage 104 are seeded are provided along the container passage 104 on the passage bottom surface 104 a of the container passage 104. ing. In the present embodiment, a recess 107 is provided concentrically with the cell seeding region 106 on the channel bottom surface 104 a of the container channel 104.

When exchanging the culture medium in the culture vessel 100, the outlet pump 10 is driven, and the old culture medium in the container passage 104 is drawn out and flows out from the container outlet 105. Along with this, a new culture medium supplied from the new solution buffer tank 6 flows into the container passage 104 from the container inlet 103. During this time, the old medium in the container passage 104 flows out of the container outlet 105 so as to be pushed out by the new medium. In this case, since the new medium and the old medium flow along the container passage 104, the new medium and the old medium can be prevented from being mixed, and the old medium can be easily replaced with the new medium.

Next, the inlet heater 5 according to this embodiment will be described with reference to FIGS. As shown in FIG. 4, the inlet heater 5 is arranged vertically in the chamber of the culture system 1.

As shown in FIGS. 4 and 5, the inlet heater 5 includes a cylindrical heater block 30 (apparatus main body) having a central axis X along the vertical direction, and a heater inlet 31 (apparatus inlet) through which a medium flows. And a heater outlet 32 (device outlet) through which the medium flows out, and a heater passage 33 (device passage) through which the medium from the heater inlet 31 toward the heater outlet 32 passes. In the present embodiment, the heater inlet 31 is provided at the upper portion 30a (preferably the upper end portion) of the heater block 30, and the heater outlet 32 is provided at the lower portion 30b (preferably the lower end portion) of the heater block 30. Yes.

The heater passage 33 is disposed on the outer periphery of the heater block 30 and is formed in a spiral shape around the central axis X of the heater block 30. As a result, in the inlet heater 5, the length of contact between the heater passage 33 and the heater block 30 can be increased, and the heat transfer area between the heater passage 33 and the heater block 30 is increased. For this reason, it is possible to sufficiently transfer heat from the heater block 30 to the medium before the medium passing through the heater passage 33 reaches the heater outlet 32. Further, since the heater inlet 31 is provided in the upper portion 30a of the heater block 30 and the heater outlet 32 is provided in the lower portion 30b of the heater block 30, the heater passage 33 formed in a spiral shape has a heater flow. It inclines downward continuously from the inlet 31 to the heater outlet 32. As a result, the medium can be prevented from remaining in the heater passage 33 when the medium is discharged, and the washing liquid can be prevented from remaining after the cleaning described later. In the present embodiment, the heater block 30 is provided with a single (one system) heater passage 33, but is not limited thereto. For example, the plurality of heater passages 33 may be provided so as to form a plurality of spirals. In this case, culture media of different systems can be heated separately.

As shown in FIGS. 5 and 6, in the present embodiment, a spiral groove 34 is provided on the outer peripheral portion of the heater block 30. The spiral groove 34 is formed in a spiral shape around the central axis X of the heater block 30. The heater passage 33 is constituted by a tube member 35, and the tube member 35 is inserted into the spiral groove 34. In this manner, the tube member 35 is spirally wound around the outer peripheral portion of the heater block 30 to form a spiral heater passage 33.

As shown in FIG. 6, the tube member 35 has a circular cross section when not inserted (restrained) in the spiral groove 34, and can be inserted into the spiral groove 34. It is suitable to have. The material used for such a tube member 35 is not particularly limited as long as it has flexibility, but for example, it is preferably formed of a flexible resin material.

The tube member 35 is pressed from the wall surface 34 a of the spiral groove 34 in a state of being inserted into the spiral groove 34. When the diameter of the tube member 35 not inserted in the spiral groove 34 is D and the width of the spiral groove 34 is W, it is preferable that D> W. In this case, in a state where the tube member 35 is inserted into the spiral groove 34, the outer surface of the tube member 35 and the wall surface 34a of the spiral groove 34 are pressed against each other, and a portion where the outer surface and the wall surface 34a contact each other without a gap is formed. The And the cross section of the tube member 35 which was circular deform | transforms so that the cross-sectional shape of the spiral groove 34 may be followed. In addition, it is suitable for the diameter D of the tube member 35 that it is a magnitude | size which is a grade which does not dent a part of the tube member 35 in the state inserted in the spiral groove 34. FIG. As an example of such a tube member 35, PFA (perfluoroalkoxy fluororesin) or PTFE (polyethylene) having a diameter of 1.6 mm (equivalent to 1/16 inch) and an inner diameter of 0.8 mm (equivalent to 1/32 inch) is used. And fluororesin tubes such as tetrafluoroethylene. In this case, the width W of the spiral groove 34 is preferably 1.55 mm. The depth H of the spiral groove 34 is preferably a depth that prevents a part of the tube member 35 from being recessed inward even when the tube member 35 is pressed from the outer peripheral side by the press tube 36 described later. For example, the thickness is preferably 1.5 mm.

As shown in FIGS. 5 and 6, a pressing tube 36 (pressing member) that presses the tube member 35 inserted into the spiral groove 34 from the outer peripheral side is provided on the outer peripheral portion of the heater block 30. The pressing tube 36 is preferably heat shrinkable. In this case, after inserting the tube member 35 into the spiral groove 34, the pressing tube 36 is attached to the heater block 30 so as to cover the entire heater passage 33. At this time, the end of the tube member 35 on the heater inlet 31 side and the end of the heater outlet 32 side protrude from the pressing tube 36. In the present embodiment, both end portions of the tube member 35 constitute the heater inlet 31 and the heater outlet 32. After the pressing tube 36 is mounted, the pressing tube 36 is contracted by heat applied from the surroundings, and is in close contact with the outer peripheral surface of the heater block 30. In this way, the tube member 35 of the heater passage 33 is pressed from the outer peripheral side by the pressing tube 36. Further, the pressing tube 36 prevents the tube member 35 inserted into the spiral groove 34 from dropping off. As an example of such a pressing tube 36, for example, a heat shrinkable tube having an inner diameter of 34 mm before heat shrinkage and an inner diameter of 26 mm after heat shrinkage may be mentioned.

As shown in FIG. 5, the heater block 30 is provided with a heater main body 37 (heating unit, for example, a cartridge heater) that heats the medium passing through the heater passage 33. The heater body 37 is attached to the heater block 30 by being inserted from below into a heater hole 38 provided in the center of the heater block 30.

The heater body 37 extends from the heater inlet 31 to the heater outlet 32 along the central axis X of the heater block 30, and it is preferable that the heat generation amount is equal over the entire length. The heater body 37 is disposed concentrically with the heater passage 33. That is, the heater main body 37 is disposed inside the spiral heater passage 33 and at the center of the heater block 30 when viewed from the direction along the central axis X. Thus, the distance between the heater main body 37 and each point of the heater passage 33 is equalized, and heat is transmitted from the heater main body 37 to each point of the heater passage 33 evenly. An example of the amount of heat generated by the heater body 37 is 2.3 W / cm ² .

The heater block 30 is preferably made of a material having good heat transfer property in order to transmit the heat generated from the heater body 37 to the heater passage 33. For example, the heater block 30 is made of a metal material such as an aluminum alloy such as A6063. It is preferred that The outer diameter of the heater block 30 is, for example, 31 mm, and the length along the central axis X is, for example, 178 mm. The number of turns of the spiral groove 34 formed in the heater block 30 is, for example, 43 turns.

1 controls the inlet pump 7 and the heater main body 37 so as to drive the heater main body 37 of the inlet heater 5 while the inlet pump 7 is being driven. Thus, the low temperature medium discharged from the medium supply source 2 is heated while passing through the heater passage 33 of the inlet heater 5, and a desired temperature (for example, about 37 ° C.) at the heater outlet 32 of the inlet heater 5. The temperature rises to

In the present embodiment, as shown in FIG. 5, the heater block 30 is provided with a temperature sensor 39 (temperature measurement unit) that measures the temperature of the heater block 30. Based on the temperature of the heater block 30 measured by the temperature sensor 39, the control unit 18 controls the heater body 37. That is, the control unit 18 drives (ON) the heater body 37 when the temperature of the heater block 30 measured by the temperature sensor 39 is less than a predetermined reference value, and when the temperature is equal to or higher than the predetermined reference value, Is stopped (OFF). Here, the predetermined reference value may be a temperature equal to the temperature of the culture medium in the culture vessel 100 (about 37 ° C.), but in order to prevent an excessive temperature rise, a lower temperature may be used. Good.

In the form shown in FIG. 5, the temperature sensor 39 is disposed in the lower part 30b of the heater block 30 (the part on the heater outlet 32 side). The temperature sensor 39 is attached to the heater block 30 by being inserted into a sensor hole 40 provided in the lower portion 30 b of the heater block 30 from below. Note that the temperature sensor 39 is not limited to being disposed in the lower portion 30b of the heater block 30, and may be disposed in the upper portion 30a of the heater block 30 as long as the medium can be prevented from overheating. Further, the temperature sensor 39 is preferably disposed between the heater passage 33 and the heater body 37 as shown in FIG. In this case, the temperature sensor 39 is disposed in a portion of the heater block 30 on the inner side of the heater passage 33 (on the heater body 37 side).

Next, the operation of the present embodiment having such a configuration will be described. First, a method of supplying a culture medium from the culture medium supply source 2 to the new solution buffer tank 6 will be described.

When supplying a new medium from the medium supply source 2 to the new solution buffer tank 6, the inlet pump 7 is driven and the heater body 37 of the inlet heater 5 is driven. As a result, a new medium stored at a low temperature is supplied to the new solution buffer tank 6 via the inlet heater 5. A new culture medium is heated to a desired temperature when passing through the inlet heater 5. The flow rate of the culture medium at this time is, for example, 15 mL / min.

More specifically, the medium supplied from the medium supply source 2 flows into the heater inlet 31 of the inlet heater 5 and passes through the heater passage 33. Since the tube member 35 constituting the heater passage 33 is inserted into the spiral groove 34 of the heater block 30, the medium passing through the heater passage 33 faces downward (heater outlet 32) in the outer periphery of the heater block 30. Flowing in a spiral. During this time, since the heater main body 37 is driven, heat generated from the heater main body 37 is transmitted to the medium passing through the heater passage 33 via the heater block 30. In particular, since the culture medium flows spirally, the heat transfer area where the culture medium can receive heat from the heater block 30 is increased. As a result, heat is efficiently transferred from the heater block 30 to the medium, and the medium is efficiently heated to a desired temperature.

Here, the temperature sensor 39 provided in the lower part 30b of the heater block 30 measures the temperature of the heater block 30 on the heater outlet 32 side. When the measured temperature is equal to or higher than a predetermined reference value, the heater main body 37 is stopped to prevent the medium from being overheated. On the other hand, when the measured temperature becomes less than the predetermined value, the heater body 37 is driven to heat the heater block 30. In this way, the culture medium passing through the heater passage 33 is heated, and the temperature of the culture medium reaching the heater outlet 32 becomes a desired temperature. In this manner, the inlet heater 5 according to the present embodiment can raise the temperature of the medium flowing from the heater inlet 31 to a desired temperature before reaching the heater outlet 32 at a low temperature.

Note that the temperature of the medium that has reached the heater outlet 32 can be affected by various parameters. Such parameters mainly include the diameter, length, and material of the heater block 30, the diameter of the tube member 35 of the heater passage 33, the number of turns, the amount of heat generated by the heater body 37, the flow rate of the culture medium, and the like. However, by sufficiently increasing the number of turns of the tube member 35, sufficient heat can be transferred from the heater block 30 to the medium passing through the heater passage 33, and the temperature of the medium is desired before reaching the heater outlet 32. The temperature can be increased to

The culture medium that has reached the heater outlet 32 flows out of the heater outlet 32 and is supplied to the buffer tank 6 for new solution. At this time, when the supply of new medium to the heater inlet 31 is stopped, the medium that has passed through the heater passage 33 of the inlet heater 5 is smoothly discharged from the heater outlet 32. That is, since the heater inlet 31 is provided in the upper part 30 a of the heater block 30 and the heater outlet 32 is provided in the lower part 30 b of the heater block 30, the heater passage 33 formed in a spiral shape has the heater inlet 31. To the heater outlet 32 continuously inclining downward. For this reason, it can suppress that a culture medium remains in the heater channel | path 33. FIG.

The medium discharged from the inlet heater 5 is supplied to the new solution buffer tank 6 and stored. In the fresh solution buffer tank 6, bubbles are removed from the stored medium. That is, since the culture medium stored in the buffer tank 6 for new solution is at a high temperature, dissolved gas and minute bubbles in the culture medium appear and expand and are likely to float. This effectively removes bubbles from the culture medium.

Next, a method for replacing the medium in the culture vessel 100 will be described.

First, the second inlet opening / closing valve 9 and the outlet opening / closing valve 12 are opened, and the medium can be circulated from the buffer tank 6 for fresh solution to the culture vessel 100. Next, the outlet pump 10 is driven, the old medium in the container passage 104 is drawn out, and flows out from the container outlet 105. As a result, the old medium is drawn out from the culture vessel 100 and discharged. At this time, a new medium stored in the buffer tank 6 for fresh solution flows into the container passage 104 via the container inlet 103 (see FIGS. 2 and 3) of the culture container 100, and the old medium in the container passage 104 The medium is drained so as to be pushed into the new medium. In this way, the container passage 104 is filled with a new medium, and the medium in the container passage 104 is exchanged.

Next, a method for cleaning the inlet heater 5 will be described.

First, the heater passage 33 is cleaned using a cleaning liquid.

In this case, first, the cleaning on-off valve 15 is opened and the second inlet on-off valve 9 is closed. As a result, the cleaning liquid from the cleaning liquid supply source 13 is supplied to the heater inlet 31 of the inlet heater 5, passes through the heater passage 33, and travels toward the heater outlet 32. The flow rate of the cleaning liquid at this time is, for example, 30 mL / min.

During this time, the heater passage 33 is cleaned by the cleaning liquid that passes therethrough. The cleaning liquid that has reached the heater outlet 32 is supplied to and stored in the new liquid buffer tank 6. When the cleaning liquid stored in the new liquid buffer tank 6 reaches a predetermined amount, the second inlet opening / closing valve 9 is opened, and the cleaning liquid stored in the new liquid buffer tank 6 flows downstream of the second inlet opening / closing valve 9. It is discharged from a solution discharge line (not shown) provided.

When the supply of the cleaning liquid to the heater inlet 31 is stopped, the cleaning liquid that has passed through the heater passage 33 of the inlet heater 5 is smoothly discharged from the heater outlet 32. That is, since the heater inlet 31 is provided in the upper part 30 a of the heater block 30 and the heater outlet 32 is provided in the lower part 30 b of the heater block 30, the heater passage 33 formed in a spiral shape has the heater inlet 31. To the heater outlet 32 continuously inclining downward. For this reason, it is possible to prevent the cleaning liquid from remaining in the heater passage 33.

However, depending on the contamination state of the heater passage 33 or the shape accuracy of the heater passage 33, a cleaning liquid may remain in the heater passage 33. Therefore, in order to further prevent the cleaning liquid from remaining in the heater passage 33, it is preferable to purge the cleaning liquid that may remain in the heater passage 33 after cleaning.

In this case, first, the gas on-off valve 17 is opened and the second inlet on-off valve 9 is closed. Thus, the compressed gas from the gas supply source 16 is supplied to the heater inlet 31 of the inlet heater 5, passes through the heater passage 33, and travels toward the heater outlet 32. The flow rate of the compressed air at this time is, for example, 30 mL / min.

During this time, the cleaning liquid remaining in the heater passage 33 is purged by the compressed gas, discharged from the heater passage 33 and supplied to the new liquid buffer tank 6. Since the space in the new solution buffer tank 6 communicates with the surrounding atmosphere through the vent portion, the compressed gas can be continuously supplied to the inlet heater 5.

Even in this case, since the spiral heater passage 33 is continuously inclined downward from the heater inlet 31 to the heater outlet 32, the cleaning liquid purged by the compressed air smoothly flows to the heater outlet 32. Can reach. For this reason, the cleaning liquid in the heater passage 33 can be more reliably discharged from the inlet heater 5. The cleaning liquid supplied to the new liquid buffer tank 6 can be discharged from a solution discharge line (not shown) provided downstream by opening the second inlet on-off valve 9.

As described above, according to the present embodiment, the heater passage 33 through which the medium from the heater inlet 31 toward the heater outlet 32 passes is formed in a spiral shape around the central axis X of the cylindrical heater block 30. Yes. As a result, the length of contact between the heater passage 33 and the heater block 30 can be increased, and the medium passing through the heater passage 33 is sufficiently heated from the heater block 30 to the medium before reaching the heater outlet 32. Can be communicated. For this reason, the culture medium can be heated while passing through the heater passage 33, and the time for heating the culture medium can be shortened.

Further, according to the present embodiment, the central axis X of the heater block 30 is along the vertical direction. Accordingly, the heater passage 33 can be continuously inclined downward, and the medium can be prevented from remaining in the heater passage 33 when the medium is discharged, and the washing liquid can be prevented from remaining after washing. . In particular, when the cleaning liquid remaining in the heater passage 33 after cleaning is purged, the cleaning liquid can smoothly reach the heater outlet 32 and the cleaning liquid remaining in the heater passage 33 can be further suppressed.

Further, according to the present embodiment, the heater inlet 31 of the inlet heater 5 is provided in the upper part 30 a of the heater block 30, and the heater outlet 32 is provided in the lower part 30 b of the heater block 30. Thereby, the heater passage 33 can be inclined downward continuously from the heater inlet 31 toward the heater outlet 32. For this reason, in order to prevent the culture medium or the cleaning liquid from remaining in the heater passage 33, the cleaning liquid or the compressed air may be introduced from the heater inlet 31, and the direction of the culture medium and the flow of the cleaning liquid and the compressed gas The direction of flow can be the same direction. In this case, it can suppress that the structure of the culture system 1 becomes complicated.

Further, according to the present embodiment, the heater passage 33 is constituted by the tube member 35 inserted into the spiral groove 34 provided in the heater block 30. Accordingly, the spiral heater passage 33 can be easily formed and the heater passage 33 can be easily replaced. In particular, according to the present embodiment, the tube member 35 of the heater passage 33 is pressed from the wall surface 34 a of the spiral groove 34. Thereby, the heat transfer resistance between the heater block 30 and the tube member 35 can be reduced, and the heat transfer efficiency from the heater block 30 to the tube member 35 can be improved. Furthermore, according to the present embodiment, the tube member 35 of the heater passage 33 is pressed from the outer peripheral side by the pressing tube 36. Thereby, the heat transfer resistance between the heater block 30 and the tube member 35 can be further reduced, and the heat transfer efficiency from the heater block 30 to the tube member 35 can be further improved.

Further, according to the present embodiment, the heater body 37 extends from the heater inlet 31 to the heater outlet 32 along the central axis X of the heater block 30 and is concentrically disposed in the heater passage 33. ing. Thus, the distance between the heater main body 37 and each point of the heater passage 33 can be equalized, and heat can be evenly transmitted from the heater main body 37 to each part of the heater passage 33. For this reason, it can prevent that the culture medium which passes the heater channel | path 33 is locally overheated, and can suppress deterioration of a culture medium.

Further, according to the present embodiment, the heater body 37 is controlled based on the temperature of the heater block 30 measured by the temperature sensor 39. Thereby, it is possible to prevent the medium passing through the heater passage 33 from being overheated and to suppress the deterioration of the medium. In particular, according to the present embodiment, since the temperature sensor 39 is disposed in the heater block 30 on the heater outlet 32 side, the heater outlet 32 on the heater passage 33 where the temperature can be increased. The temperature of the culture medium in this part can be measured, and the heater body 37 can be controlled based on this temperature. For this reason, it can prevent further that a culture medium is overheated, and can suppress further deterioration of a culture medium. Further, since the temperature sensor 39 is disposed between the heater passage 33 and the heater main body 37, the temperature sensor 39 can measure the temperature in the heater block 30 on the inner side (the heater main body 37 side) of the heater passage 33. The heater body 37 can be controlled based on this temperature. For this reason, it can prevent further that a culture medium is overheated, and can suppress further deterioration of a culture medium.

Further, according to the present embodiment, the medium heated by the inlet heater 5 is stored in the buffer tank 6 for new solution. Thereby, bubbles can be removed from the stored medium. That is, since the culture medium stored in the buffer tank 6 for new solution is at a high temperature, dissolved gas and minute bubbles in the culture medium appear and expand, and the bubbles can be efficiently removed.

Furthermore, according to the present embodiment, the inlet pump 7 is provided between the culture medium supply source 2 and the inlet heater 5. Thereby, it is possible to suppress the formation of bubbles in the culture medium in the heater passage 33 due to an increase in the pressure of the culture medium passing through the heater passage 33 of the inlet heater 5. For this reason, heat can be efficiently transmitted from the heater block 30 to the culture medium.

In addition, in this Embodiment mentioned above, while the heater inflow port 31 was provided in the upper part 30a of the heater block 30, the heater outflow port 32 was provided in the lower part 30b of the heater block 30 was demonstrated. However, the present invention is not limited to this, and the heater inlet 31 may be provided in the lower part 30 b of the heater block 30, and the heater outlet 32 may be provided in the upper part 30 a of the heater block 30. In this case, in order to prevent the culture medium or the cleaning liquid from remaining in the heater passage 33, the cleaning liquid or the compressed air may be introduced from the heater outlet 32.

In the above-described embodiment, the example in which the tube member 35 of the heater passage 33 is pressed from the wall surface 34a of the spiral groove 34 provided in the heater block 30 has been described. However, the present invention is not limited to this, and the tube member 35 may not be pressed from the wall surface 34 a of the spiral groove 34 as long as heat transfer efficiency from the heater block 30 to the tube member 35 can be ensured.

In the above-described embodiment, the example in which the tube member 35 of the heater passage 33 is pressed from the outer peripheral side by the pressing tube 36 has been described. However, the present invention is not limited to this, and if the heat transfer efficiency from the heater block 30 to the tube member 35 can be ensured and the tube member 35 inserted into the spiral groove 34 can be prevented from falling off, the pressure can be reduced. The tube 36 may not be provided. Further, the pressing tube 36 may not have heat shrinkability, and may be formed using, for example, a material having elasticity. In this case, the inner diameter of the pressing tube 36 may be smaller than the outer diameter of the heater block 30 and may be attached to the outer peripheral surface of the heater block 30 while being elastically deformed so as to expand.

Furthermore, in the above-described embodiment, the new solution buffer tank 6 may have a heater (not shown) for heating the stored medium. In this case, the stored medium can be maintained at a desired temperature. Further, even when the temperature of the medium discharged from the inlet heater 5 is not raised to a desired temperature, the temperature is raised to the desired temperature while being stored in the new solution buffer tank 6. Can do. Even in this case, since the low temperature medium can be brought close to a desired temperature by the inlet heater 5, it is possible to contribute to shortening the time for heating the medium.

The present invention is not limited to the above-described embodiments and modification examples as they are, and can be embodied by modifying the constituent elements without departing from the scope of the invention in the implementation stage. Further, various inventions can be formed by appropriately combining a plurality of constituent elements disclosed in the above-described embodiments and modifications. You may delete a some component from all the components shown by embodiment and a modification. Furthermore, constituent elements over different embodiments and modifications may be combined as appropriate.

Claims

A medium heating apparatus for heating a medium for cell culture supplied from a medium supply source to a culture container,
A cylindrical device body having a central axis along the vertical direction;
A device inlet provided in one of an upper part and a lower part of the apparatus main body, into which the culture medium flows;
A device outlet provided on the other of the upper part and the lower part of the apparatus main body, through which the medium flows out;
A device passage through which the culture medium passes from the device inlet to the device outlet;
A heating unit for heating the culture medium passing through the device passage,
The apparatus passage is a medium heating apparatus formed in a spiral shape around the central axis of the apparatus main body.
The apparatus inlet is provided on the upper part of the apparatus body,
The culture apparatus according to claim 1, wherein the apparatus outlet is provided at a lower portion of the apparatus main body.
A spiral groove is provided on the outer periphery of the apparatus body,
The spiral groove is formed in a spiral shape around the central axis of the device body,
The culture apparatus according to claim 1, wherein the device passage is configured by a tube member inserted into the spiral groove.
The culture medium heating apparatus according to claim 3, wherein the tube member is pressed from a wall surface of the spiral groove.
The culture medium heating apparatus according to claim 3 or 4, further comprising a pressing member that presses the tube member from the outer peripheral side.
6. The heating unit according to claim 1, wherein the heating unit extends from the apparatus inlet to the apparatus outlet along the central axis of the apparatus main body and is concentrically disposed in the apparatus passage. The culture medium heating apparatus according to claim 1.
The culture medium heating apparatus according to any one of claims 1 to 6, further comprising a temperature measurement unit that measures the temperature of the apparatus main body.
The culture medium heating apparatus according to claim 7, wherein the temperature measuring unit is arranged in a part of the apparatus main body on the apparatus outlet side.
The culture medium heating device according to claim 7 or 8, wherein the temperature measurement unit is disposed between the device passage and the heating unit.
A medium source for supplying new medium;
A container holding unit capable of holding a culture container;
The medium heating apparatus according to claim 1, wherein the medium heating apparatus is provided between the medium supply source and the container holding unit, and heats the medium supplied from the medium supply source to the culture container. Culture system.
The culture system according to claim 10, further comprising a buffer tank provided between the culture medium heating device and the container holding unit and storing the culture medium heated by the culture medium heating device.
The culture system according to claim 10 or 11, further comprising a medium supply pump that is provided between the medium supply source and the medium heating apparatus and supplies the medium from the medium supply source to the medium heating apparatus.
A control unit for controlling the heating unit of the medium supply pump and the medium heating device;
The culture medium heating device has a temperature measurement unit that measures the temperature of the device main body,
The said control part controls the said culture medium supply pump and the said heating part based on the temperature of the said apparatus main body measured by the said temperature measurement part, while driving the said culture medium supply pump. Culture system.
The culture according to any one of claims 10 to 13, further comprising a cleaning liquid supply source for supplying a cleaning liquid to the apparatus inlet or the apparatus outlet provided in an upper part of the apparatus main body of the culture medium heating apparatus. system.
The culture according to claim 14, further comprising a gas supply source for supplying a gas capable of purging the cleaning liquid to the apparatus inlet or the apparatus outlet provided in an upper part of the apparatus main body of the culture medium heating apparatus. system.