Cell culture system, culture tank, and culture tank connector

Abstract

The present invention provides a cell culture system which enables a simplification of the overall configuration thereof and which makes it possible to reduce the processing load on a control device. Provided is a cell culture system (S1) comprising: a plurality of culture tanks (100, 200, 300) which are connected in series or in parallel via tubular members (60-63) and which store a culture solution (50); a pump (68), the number of which is less than the number of culture tanks, and which causes the culture solution to flow between the plurality of culture tanks; and a control device (70) which controls the pump, wherein the plurality of culture tanks each have a pair of connection ports (3a, 3c) on side surfaces at a lower part thereof, and the tubular members are connected to the pairs of connection ports via connectors (10).

Description

Cell culture system, culture vessel, and connector for culture vessel

[0001] The present invention relates to a cell culture system, a culture vessel, and a connector for a culture vessel.

[0002] In recent years, countries around the world have been developing truly sustainable food production systems in order to achieve the Sustainable Development Goals (SDGs), and in particular, development of "cellular foods" has been expanding. Furthermore, as background art in this technical field, for example, Patent Document 1 describes a cell culture system in which multiple culture vessels are connected and a culture medium is circulated between the multiple culture vessels.

[0003] International Publication No. 2017 / 191691

[0004] When connecting multiple culture vessels with tubular members such as hoses, a structure is generally used in which the culture medium is supplied and discharged from the top of the culture vessels. However, when this structure is applied to the cell culture system described in Patent Document 1, for example, a pump is required for each culture vessel. This increases the overall size of the cell culture system, and also poses the problem of a large processing load on the control device due to the need to simultaneously control multiple pumps.

[0005] The present invention has been made in view of the above circumstances, and its main object is to provide a cell culture system that can simplify the overall configuration and reduce the processing load on the control device. Another object of the present invention is to provide a culture vessel and a culture vessel connector that are suitable for the cell culture system.

[0006] In order to achieve the above object, a first aspect of the present invention is a cell culture system comprising a plurality of culture tanks connected in series or in parallel via tubular members and storing a culture medium, a pump whose number is less than the number of the culture tanks and for flowing the culture medium among the plurality of culture tanks, and a control device for controlling the pump, wherein each of the plurality of culture tanks has a pair of connection ports on its side and bottom, and the tubular member is connected to each of the pair of connection ports via a connector.

[0007] In order to achieve the above object, a second aspect of the present invention is a culture tank for storing a culture medium, comprising a pair of connection ports provided on a side surface and a lower portion thereof, and connectors provided on each of the pair of connection ports for connecting a tubular member, the connectors including a connector body that is inserted into the connection ports and has a flow path formed therein through which the culture medium flows, a pair of gaskets that are fitted onto the connector body, and a nut that is fitted onto the connector body and holds the pair of gaskets, the connector body having a flange portion provided at one end and having an outer diameter larger than the connection ports, a fitting portion provided at the other end and fitting with the tubular member, and a threaded portion provided between the flange portion and the fitting portion and threadedly engaging with the nut, the connector body being characterized in that the flange portion is located inside the culture tank and the fitting portion is located outside the culture tank, one of the pair of gaskets is disposed between the inner wall of the culture tank and the flange portion, and the other is disposed between the outer wall of the culture tank and the nut.

[0008] In order to achieve the above object, a third aspect of the present invention is a connector for a culture tank that is attached to a connection port of a culture tank that stores a culture medium and that connects a tubular member, the connector comprising: a connector body that is inserted into the connection port and has a flow path through which the culture medium flows; a pair of gaskets that are fitted onto the connector body; and a nut that is fitted onto the connector body and that holds the pair of gaskets. The connector body has a flange portion that is provided at one end and has an outer diameter larger than the connection port; a fitting portion that is provided at the other end and that fits with the tubular member; and a threaded portion that is provided between the flange portion and the fitting portion and that screws onto the nut. The connector body is inserted into the connection port so that the flange portion is located inside the culture tank and the fitting portion is located outside the culture tank. One of the pair of gaskets is disposed between the inner wall of the culture tank and the flange portion, and the other is disposed between the outer wall of the culture tank and the nut.

[0009] According to the present invention, it is possible to provide a cell culture system that can simplify the overall configuration and reduce the processing load of the control device. Furthermore, according to the present invention, it is possible to provide a culture vessel and a culture vessel connector that are suitable for the cell culture system. Note that problems, configurations, and effects other than those described above will become clear from the description of the following embodiments.

[0010] Fig. 1 is an overall configuration diagram schematically showing a cell culture system according to a first embodiment. Fig. 2 is a configuration diagram of the cell culture system shown in Fig. 1 as seen from above. Fig. 3 is a perspective view schematically showing a culture vessel. Fig. 4 is an exploded view of a connector. Fig. 5 is a partially enlarged cross-sectional view showing a state in which the connector is inserted into the connection port of the culture vessel. Fig. 6 is a configuration diagram of a cell culture system according to a second embodiment as seen from above.

[0011] Hereinafter, an embodiment of the present invention will be described with reference to the drawings, taking the case of culturing animal cells as an example.

[0012] (First embodiment) A cell culture system S1 according to a first embodiment of the present invention will be described. Fig. 1 is a schematic diagram of the overall configuration of the cell culture system S1 according to the first embodiment, and Fig. 2 is a configuration diagram of the cell culture system S1 shown in Fig. 1 as seen from above. Note that in Fig. 2, a pump 68 and a controller 70 are not shown.

[0013] 1 and 2, the cell culture system S1 according to this embodiment has, for example, three culture vessels 100, 200, and 300 arranged in series along the flow of the culture solution, and the culture vessels 100, 200, and 300 are connected in sequence via hoses 60 to 63, which are an example of tubular members, to form a ring. The number of culture vessels is not limited to three. Furthermore, pipes, tubes, etc. may be used instead of the hoses 60 to 64.

[0014] The culture tanks 100, 200, and 300 are vessels made of, for example, stainless steel or glass, and contain a culture solution 50 therein.

[0015] In this embodiment, different animal cells are supplied to the culture vessels 100, 200, and 300, but the same cells may be supplied to some or all of the culture vessels 100, 200, and 300. The animal cells supplied to the culture vessels 100, 200, and 300 attach to the scaffold 40 and are cultured to produce a cellular food product. In addition, the cells secrete useful components into the culture medium during the culture process, which can be used as the culture supernatant.

[0016] The scaffold 40 is a sheet or three-dimensional structure with a porous or gel-like structure for attachment of animal cells. The material of the scaffold 40 is not particularly limited, but examples include resins such as PET and PP, and materials derived from plants or animals. In this embodiment, the scaffold 40 is an edible scaffold derived from plants or animals. The scaffold 40 includes, for example, proteins and polysaccharides. Here, "edible" refers to a composition that can be safely ingested into the body. Cellular food is produced when animal cells attach and grow on the scaffold 40 as a substrate.

[0017] Examples of animal cells that adhere to the scaffold 40, i.e., animal cells that are supplied to the culture vessels 100, 200, and 300, include muscle-derived cells, skin-derived cells, intestinal cells, heart-derived cells, brain-derived cells, stomach-derived cells, embryonic membrane-derived cells, liver-derived cells, kidney-derived cells, and lung-derived cells. Among these, by culturing different types of cells in the culture vessels 100, 200, and 300, a more useful culture supernatant can be produced.

[0018] Furthermore, the cell culture system S1 includes one pump 68. The pump 68 is disposed between the hose 60 and the hose 63, and is driven by a motor (not shown). When the pump 68 is driven, the culture solution 50 circulates between the culture tanks 100 and 300 in the direction of the arrow F in the figure. As the rotation speed of the motor changes, the flow rate of the pump 68 also changes. That is, the circulation flow rate of the culture solution 50 flowing through the cell culture system S1 changes. The number of pumps 68 is not limited to one. It is sufficient if the number is less than the number of culture tanks.

[0019] The controller 70 is connected to the pump 68 (more specifically, the motor that drives the pump 68) via electrical wiring E1. Furthermore, the controller 70 is also connected to various sensors (temperature sensor, flow rate sensor, pH detection sensor, etc.) not shown via electrical wiring, and is capable of monitoring the states of the culture tanks 100, 200, 300 based on sensor data input from the various sensors. Note that, although the controller 70 is configured separately from the pump 68 in FIG. 1, the controller 70 and the pump 68 may be configured as an integrated unit.

[0020] The controller 70 includes a processing unit having a CPU, ROM and RAM as storage devices, and other peripheral circuits. A control program is stored in the ROM, and the CPU reads the control program and executes various processes. The controller 70 controls, for example, the flow rate of the pump 68 based on various input information.

[0021] Next, a detailed description will be given of the structures of the culture tanks 100, 200, and 300. Since the culture tank 100 has the same configuration as the culture tanks 200 and 300, the following description will focus on the culture tank 100, and descriptions of the culture tanks 200 and 300 will be omitted.

[0022] FIG. 3 is a perspective view that schematically illustrates a culture tank 100. As shown in FIG. 3, the culture tank 100 includes a vessel body 1 and a lid 2 that closes an opening provided at the top of the vessel body 1. The vessel body 1 has a hollow rectangular parallelepiped shape, and a pair of connection ports 3a, 3c are provided on opposing side surfaces 1a and 1c of four side surfaces 1a to 1d. That is, the connection ports 3a and 3c are arranged to face each other. The connection port 3a is an inlet for the culture solution 50, and the connection port 3c is an outlet for the culture solution 50 (see FIG. 2).

[0023] Connection port 3a is provided at a height of position H1 from the bottom surface 1e of container body 1. Similarly, connection port 3c is provided at a height of position H2 from the bottom surface 1e of container body 1. In this embodiment, positions H1 and H2 are at the same height. Here, positions H1 and H2 are preferably approximately ⅓ or less of the height of container body 1 from the bottom surface 1e of container body 1. By providing connection ports 3a and 3c on the side and bottom of container body 1 in this way, smooth circulation of culture solution 50 is ensured even with a single pump 68.

[0024] The lid 2 closes the opening of the container body 1 to seal the inside of the container body 1. Hoses 5 pass through the lid 2, and nutrients such as oxygen are supplied through the hoses 5.

[0025] A connector 10 is inserted into the connection ports 3a, 3c, and hoses 60-63 are attached via the connector 10 (see FIG. 2). Details of the connector 10 will be described with reference to FIG. 4, which is an exploded view of the connector 10. As shown in FIG. 4, the connector 10 is configured to include a connector body 11, a pair of packings 21, 22, and a nut 31.

[0026] The connector body 11 has a flange portion 12 provided at one end, a fitting portion 14 provided at the other end, and a male thread portion (screw portion) 13 provided between the flange portion 12 and the fitting portion 14. The connector body 11 has a flow path 15 extending axially therein. The culture solution 50 flows through this flow path 15. The flange portion 12 is formed with an outer diameter larger than the connection ports 3a, 3c. The fitting portion 14 is inserted into and fitted with the hoses 60-63. Therefore, it is formed with an outer diameter approximately equal to the inner diameter of the hoses 60-63. The male thread portion 13 threads into a female thread portion 32 of a nut 31, which will be described later.

[0027] The pair of packings 21, 22 are made of, for example, silicone resin. The pair of packings 21, 22 are each fitted onto the connector body 11. Therefore, the inner diameters of the holes 21a, 22a of the packings 21, 22 are slightly larger than the outer diameter of the male thread portion 13 of the connector body 11.

[0028] The nut 31 is fitted onto the connector body 11. The nut 31 has a pair of protrusions 33 on its periphery. The protrusions 33 are intended to engage the fingers of an operator when rotating the nut 31. The female thread portion 32 of the nut 31 is threadedly engaged with the male thread portion 13 of the connector body 11, thereby holding the pair of gaskets 21, 22 between the nut 31 and the flange portion 12.

[0029] Next, a method for attaching the connector 10 will be described. Fig. 5 is a partially enlarged cross-sectional view showing the connector 10 inserted into the connection port 3a of the culture tank 100. As shown in Fig. 5, when the connector body 11 is inserted from the inside of the connection port 3a (inside the vessel body 1) to the outside with the packing 21 fitted over the connector body 11, the flange portion 12 abuts against the side surface 1a of the vessel body 1 via the packing 21. In other words, the packing 21 is sandwiched between the flange portion 12 and the side surface 1a of the vessel body 1.

[0030] Next, the packing 22 is fitted onto the fitting portion 14 of the connector body 11 exposed to the outside of the container body 1 from the connection port 3a, and then the nut 31 is fitted onto the connector body 11 and tightened. Then, the pair of packings 21, 22 seal the connection port 3a.

[0031] In this state, the hose 60 is inserted into and fixed to the fitting portion 14 of the connector body 11. In this way, the hose 60 and the container body 1 are connected via the flow path 15 of the connector 10. That is, the culture solution 50 can flow from the hose 60 into the container body 1. The connector 10 is attached to the connection port 3c in the same manner.

[0032] According to this embodiment configured as above, the following advantageous effects can be achieved.

[0033] For example, because the hoses 60 to 63 are connected to the connection ports 3a and 3c provided at the bottom of the culture tanks 100, 200, and 300, there is no need to pump the culture solution 50 up to a position higher than the culture tanks when circulating the culture solution 50. Therefore, the culture solution 50 can be circulated with fewer pumps than the number of culture tanks (three in this embodiment), specifically with one pump 68. This simplifies the overall configuration of the cell culture system S1.

[0034] Furthermore, since the controller 70 only needs to control one pump 68, the burden of control processing is reduced compared to when multiple pumps are controlled. Therefore, this embodiment also contributes to the achievement of the Sustainable Development Goals (SDGs). Furthermore, because the configuration of the culture vessels 100, 200, and 300 themselves is simple, they can be easily incorporated into various devices to build a cell culture system, and are highly scalable.

[0035] In addition, because the connection ports 3a and 3c are located opposite each other, it is easy to arrange the culture vessels 100, 200, and 300 in a line, and it is also easy to route the hoses 60 to 63. Furthermore, because the height positions H1 and H2 of the connection ports 3a and 3c are the same, the hoses 60 to 63 can be routed without having to worry about the orientation of the culture vessels 100, 300, and 300. This makes it possible to improve the efficiency of the assembly work of the cell culture system S1.

[0036] Furthermore, because the connector 10 is configured to be inserted into the connection ports 3a, 3c and connect the hoses 60-63 to the vessel body 1 via the connector 10, the connection process for the culture tanks 100, 200, and 300 is simple. In particular, the flange portion 12 of the connector 10 is pressed against the inner wall of the vessel body 1 via the packing 21, and then the nut 31 is tightened so that the packing 22 is pressed against the outer wall of the vessel body 1, completing the attachment of the connector 10 to the connection ports 3a, 3c. At this time, the packings 21, 22 seal the connection ports 3a, 3c, eliminating the risk of leakage of the culture medium 50. The hoses 60-63 are then fitted into the fitting portion 14, completing the routing of the hoses 60-63. In this way, the cell culture system S1 can be easily constructed using the connector 10. Furthermore, if pipes or tubes are used instead of the hoses 60-63, simply changing the shape of the fitting portion 14 of the connector body 11 is sufficient, resulting in high versatility.

[0037] Second Embodiment Next, a cell culture system S2 according to a second embodiment of the present invention will be described. Fig. 6 is a configuration diagram of the cell culture system S2 according to the second embodiment as seen from above. Note that in Fig. 6, the pump 68 and the controller 70 are not shown.

[0038] As shown in Figure 6, the second embodiment is characterized in that the positions of the connection ports provided in the culture tanks 100, 200, and 300 are different from those in the first embodiment. Specifically, in the culture tank 100, connection ports 3a and 3b are provided on the side surfaces 1a and 1b (see Figure 3) of the vessel body 1. That is, the connection ports 3a and 3b are not provided in opposing positions, but the connection port 3b is provided at a 90-degree angle from the connection port 3a in a top view. The culture tanks 200 and 300 are configured in a similar manner. Note that the height positions of the connection ports 3a, 3b, 3c, and 3d are all the same.

[0039] According to the second embodiment configured as described above, it is possible to achieve the same effects as those of the first embodiment. Moreover, since the culture tanks 100, 200, and 300 can be arranged without lining up in a line, there is an advantage in that the degree of freedom in layout is increased.

[0040] The present invention is not limited to the above-described embodiments, and various modifications are possible within the scope of the gist of the present invention. The present invention covers all technical matters included in the technical ideas described in the claims. The above-described embodiments are preferred examples, but a person skilled in the art can realize various alternatives, modifications, variations, or improvements from the contents disclosed in this specification, and these are included in the technical scope described in the appended claims.

[0041] For example, the present invention can also be applied as an apparatus for culturing cells other than those derived from animals (such as cells derived from plants).

[0042] Furthermore, instead of connecting the culture vessels 100, 200, and 300 in series to form a cell culture system, these culture vessels may be connected in parallel to form a cell culture system.

[0043] The connection ports 3a and 3c may also be configured to have different heights. In this case, it is preferable to make the connection port 3a, which is the inlet side of the culture solution 50, higher than the connection port 3c, which is the outlet side (i.e., H1 > H2). In this way, the head difference increases the convection of the culture solution 50, and the culture solution 50 is distributed more uniformly throughout the culture tank 100, thereby supplying more nutrients and oxygen to the cells in the culture tank 100.

[0044] Furthermore, the connection ports 3a and 3c may be positioned at different widthwise positions. In this case, for example, in Figure 3, the connection port 3a, which is the inlet side of the culture solution 50, is provided on the left side of the culture tank 100, and the connection port 3c, which is the outlet side, is provided on the right side of the culture tank 100, thereby increasing the distance between the outlet and the inlet. This increases the convection of the culture solution 50, and the culture solution 50 is distributed more uniformly throughout the culture tank 100, allowing more nutrients and oxygen to be supplied to the cells in the culture tank 100.

[0045] Furthermore, the connector 10 can be provided on the top of the container body 1 or on the lid 2 and used for gas exchange in the container body 1 .

[0046] Here, the container body 1 is not limited to a hollow rectangular parallelepiped shape. For example, a cylindrical container body or a container body of other shapes may be applied to the present invention. In this case, if it is desired to arrange the pair of connection ports in opposing positions, they may be arranged so as to be approximately opposing to each other in accordance with the shape of the container.

[0047] Furthermore, the material of the pair of packings 21 and 22 is not limited to silicone. Any packing material can be used as long as it can ensure the airtightness of the container body 1. Furthermore, in addition to the pair of packings 21 and 22, metal washers can also be used.

[0048] DESCRIPTION OF SYMBOLS 1 Container body 1a to 1d Side surface 2 Lid body 3a to 3d Connection port 5 Hoses 10 Connector 11 Connector body 12 Flange portion 13 Male thread portion (screw portion) 14 Fitting portion 15 Flow path 21, 22 Packing 31 Nut 32 Female thread portion 33 Protrusion 50 Culture solution 60 to 63 Hoses (tubular members) 68 Pump 70 Controller (control device) 100, 200, 300 Culture tank S1, S2 Cell culture system

Claims

1. A cell culture system comprising: a plurality of culture tanks connected in series or parallel via tubular members and for storing culture medium; a pump, the number of which is less than the number of the culture tanks, for flowing the culture medium among the plurality of culture tanks; and a control device for controlling the pump, wherein each of the plurality of culture tanks has a pair of connection ports on a side surface and bottom thereof, and the tubular member is connected to each of the pair of connection ports via a connector.

2. The cell culture system according to claim 1, wherein the pair of connection ports are arranged at opposing positions.

3. A cell culture system according to claim 1 or 2, characterized in that the pair of connection ports are arranged at the same height position.

4. A cell culture system according to claim 1, wherein the connector includes: a connector body that is inserted into the connection port and has a flow path formed therein through which the culture medium flows; a pair of gaskets that are fitted onto the connector body; and a nut that is fitted onto the connector body and holds the pair of gaskets; the connector body has a flange portion that is provided at one end and has an outer diameter larger than the connection port, a fitting portion that is provided at the other end and fits with the tubular member, and a threaded portion that is provided between the flange portion and the fitting portion and screws into the nut; the connector body is inserted into the connection port so that the flange portion is located inside the culture tank and the fitting portion is located outside the culture tank; and one of the pair of gaskets is positioned between the inner wall of the culture tank and the flange portion, and the other is positioned between the outer wall of the culture tank and the nut.

5. A culture tank for storing a culture medium, comprising: a pair of connection ports provided on a side surface and a lower portion; and connectors provided on each of the pair of connection ports for connecting a tubular member, the connectors including: a connector body inserted into the connection ports and having a flow path formed therein through which the culture medium flows; a pair of gaskets fitted onto the connector body; and a nut fitted onto the connector body for holding the pair of gaskets, the connector body having: a flange portion provided at one end and formed with an outer diameter larger than the connection ports; a fitting portion provided at the other end for fitting with the tubular member; and a threaded portion provided between the flange portion and the fitting portion for screwing with the nut, the connector body being inserted into the connection port such that the flange portion is located inside the culture tank and the fitting portion is located outside the culture tank, one of the pair of gaskets is disposed between an inner wall of the culture tank and the flange portion, and the other is disposed between an outer wall of the culture tank and the nut.

6. A culture tank connector for attaching to a connection port of a culture tank that stores culture medium and for connecting a tubular member, comprising: a connector body that is inserted into the connection port and has a flow path formed therein through which the culture medium flows; a pair of gaskets that are fitted onto the connector body; and a nut that is fitted onto the connector body and holds the pair of gaskets, wherein the connector body has: a flange portion that is provided at one end and has an outer diameter larger than the connection port; a fitting portion that is provided at the other end and fits with the tubular member; and a threaded portion that is provided between the flange portion and the fitting portion and screws into the nut, wherein the connector body is inserted into the connection port such that the flange portion is located inside the culture tank and the fitting portion is located outside the culture tank, and one of the pair of gaskets is located between an inner wall of the culture tank and the flange portion, and the other is located between an outer wall of the culture tank and the nut.

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