Cell production cartridge and system

Abstract

To secure safety without necessitating a facility investment associated with the risk of liquid leakage.SOLUTION: According to one embodiment, a cell production cartridge includes a closed-system flow channel and a closed-system container for accommodating the flow channel. The container includes an energy transmission part for receiving, from outside the container, energy for sending a liquid within the flow channel. The energy transmission part is located at a boundary between the inside and the outside of the container.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The embodiments disclosed in the specification and drawings relate to a cartridge for cell production. [Background technology]

[0002] The cell production process includes, for example, a step of extracting cells from a donor's tissue and a step of culturing the cells. These steps are performed by combining large equipment such as a centrifuge with manual techniques by engineers. However, the preparation and maintenance of large equipment is very expensive. In addition, the work in the series of steps is complex and delicate, and the quality of the cells varies depending on the skill of the individual involved in the manual work.

[0003] Therefore, in order to produce high-quality cells at low cost, it is necessary to automate the cell production process using equipment. However, if the process is automated using equipment, there is a possibility that unexpected pressure may be applied when cells are sent to the flow path of the piping, causing the piping to come off and the liquid to leak out. For this reason, it is necessary to create an environment that ensures safety, such as a safety cabinet or a laboratory that complies with Biosafety Level 2 (BSL-2).

[0004] However, in order to construct a safety cabinet or BSL-2 compatible environment, a large amount of capital investment is required in addition to the equipment, making it difficult to manufacture at low cost. Therefore, it is desirable to ensure safety without making capital investments that are associated with the possibility of liquid leakage. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2016-118681 A Summary of the Invention [Problem to be solved by the invention]

[0006] One of the problems to be solved by the embodiments disclosed in this specification and the drawings is to ensure safety without making capital investments associated with the possibility of liquid leakage. However, the problems to be solved by the embodiments disclosed in this specification and the drawings are not limited to the above problem. Problems corresponding to the effects of each configuration shown in the embodiments described below can also be positioned as other problems. [Means for solving the problem]

[0007] The cell preparation cartridge according to the embodiment includes a flow path and a container that houses the flow path. The container includes an energy transmission unit that receives energy from outside the container for sending a fluid through the flow path. The energy transmission unit is located at a boundary that separates the inside and outside of the container. [Brief description of the drawings]

[0008] [Figure 1] FIG. 1 is a perspective view that illustrates an example of a cell production cartridge according to one embodiment and its peripheral configuration. [Diagram 2] FIG. 2 is a perspective view that illustrates a typical appearance of the cell production cartridge of FIG. [Diagram 3] FIG. 3 is a cross-sectional view taken along line III-III in FIG. [Figure 4] FIG. 4 is a schematic diagram for explaining the function of the cell production cartridge of FIG. [Diagram 5] FIG. 5 is a schematic diagram for explaining a diaphragm through which the cell production cartridge receives energy from the liquid delivery mechanism in FIG. [Figure 6] FIG. 6 is a schematic diagram for explaining a balloon through which the cell production cartridge receives energy from the liquid delivery mechanism in FIG. [Figure 7] FIG. 7 is a cross-sectional view that illustrates a state in which the pin member in the cell production cartridge of FIG. 2 opens the flow path. [Figure 8] FIG. 8 is a cross-sectional view that illustrates a state in which a pin member in the cell production cartridge of FIG. 2 blocks a flow path. [Figure 9] FIG. 9 is a flowchart illustrating an example of an operation in an embodiment. [Figure 10] FIG. 10 is a schematic diagram for explaining an example of the operation in one embodiment. [Figure 11] FIG. 11 is a schematic diagram for explaining an example of the operation in one embodiment. [Figure 12] FIG. 12 is a schematic diagram for explaining an example of the operation in one embodiment. [Figure 13] FIG. 13 is a schematic diagram for explaining an example of the operation in one embodiment. [Figure 14] FIG. 14 is a schematic diagram for explaining an example of the operation in one embodiment. [Figure 15] FIG. 15 is a schematic diagram for explaining a bellows which is a modified example of one embodiment. [Figure 16] FIG. 16 is a flowchart for explaining the operation in another modified example of the embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] Hereinafter, a cell production cartridge according to one embodiment will be described with reference to the drawings. Note that the following description will be given by taking as an example a case where the cell production cartridge is used for producing induced pluripotent stem (iPS) cells. However, the cell production cartridge can be used for producing any cell, not limited to iPS cells.

[0010] FIG. 1 is a perspective view showing an example of a cell production cartridge according to an embodiment and its peripheral configuration, and FIG. 2 is a perspective view showing an external appearance of the cell production cartridge of FIG. 1. FIG. 3 is a cross-sectional view taken along the line III-III in FIG. 2. The cell production cartridge 1 is disposed, for example, in the vicinity of a liquid delivery mechanism 4 and is detachably held in a culture device 5. The cell production cartridge 1 includes a closed flow path and a closed container for accommodating the flow path. The closed flow path has a first closed structure formed by connecting a tube, a reservoir, or the like as a flow path for transporting a liquid containing cells. The closed container has a second closed structure disposed outside the flow path of the first closed structure so that the liquid does not leak out of the cell production cartridge 1. That is, the cell production cartridge 1 has a double closed structure consisting of the first closed structure and the second closed structure. Note that the closed flow path may be connected to the outside of the flow path or the outside of the closed container via a sterile filter as long as sterility is ensured by the sterile filter. In this case, the sterile filter is preferably hydrophobic. The container of the cell preparation cartridge 1 is made of resin or plastic, and includes a container body 2 having a recess formed therein for holding the flow path, and a lid 3 attached to the container body 2 so as to cover the recess. The container body 2 and the lid 3 are hard. Here, being hard means that the elastic modulus is 10 times or more the elastic modulus of a soft part such as a tube 20 or a bag described later. The container body 2 and the lid 3 are made of, for example, resin or plastic. The resin is, for example, polycarbonate (PC) or polystyrene (PS). At least one of the container body 2 and the lid 3 is preferably light-transmitting. For example, when at least one of the container body 2 and the lid 3 has visible light transmittance, it is preferable from the viewpoint of easily observing the cell preparation process. FIG. 2 shows the appearance when the container body 2 is opaque and the lid 3 is transparent. The recess of the container body 2 includes holes 11H, 12H, 14H-17H, and 19H that accommodate the ends of the closed system flow path, holes 13H and 18H that accommodate the intermediate portion of the closed system flow path, and a groove 20T that is formed to connect to the holes 11H-19H and that accommodates the intermediate portion of the closed system flow path.The hole 3a of the lid 3 is located above the hole 19H of the recess of the container body 2, and exposes the balloon 19 accommodated in the hole 19H. The lid 3 is provided with a diaphragm 3D at a position covering the holes 11H, 12H, 14H, and 15H of the recess of the container body 2. In other words, the lid 3 is provided with a diaphragm 3D at a position covering a storage part of the flow path. The diaphragm 3D and the balloon 19 are an example of an energy transfer part that receives energy from the outside of the container for sending a fluid in a closed flow path. The diaphragm 3D and the balloon 19 are located at the boundary separating the inside and the outside of the container. The pin members 31 to 38 and the handle 39 of the cell production cartridge 1 penetrate the lid 3 and are slidably held in a part of the groove 20T of the container body 2. The layout of the recess corresponding to the number of holes 11H to 19H and the shape of the groove 20T can be any layout depending on the cell production process. However, when both the liquid delivery mechanism 4 and the culture device 5 are attached to the cell production cartridge 1, the area related to the liquid delivery mechanism 4 (holes 11H, 12H, 14H, 15H, 19H) and the area related to the culture device 5 (hole 17H) are laid out apart to avoid interference. On the other hand, when the liquid delivery mechanism 4 and the culture device 5 are attached to the cell production cartridge 1 one by one, the area related to the liquid delivery mechanism 4 (holes 11H, 12H, 14H, 15H, 19H) and the area related to the culture device 5 (hole 17H) may be laid out close to each other. Similarly, any layout can be used for the hole 3a and the diaphragm 3D of the lid 3, and the pin members 31 to 38 and the handle 39 of the cell production cartridge 1. The container of the cell production cartridge 1 may communicate with the inside and outside of the container via the sterile filter. In this case, the sterile filter is preferably hydrophobic.

[0011] The liquid delivery mechanism 4 includes a first mechanism 41 that applies energy to the diaphragm 3D and a second mechanism 42 that applies energy to the balloon 19. The first mechanism 41 includes a first arm 41a whose tip is located above the diaphragm 3D, and a rod-shaped member 41b that is held and controlled by the tip of the first arm 41a and presses the piston via the diaphragm 3D. The number of first mechanisms 41 is the same as the number of diaphragms 3D, and can be changed appropriately according to the number of diaphragms 3D. The second mechanism 42 includes a second arm 42a whose tip is located above the balloon 19, and a columnar member 42b that is held and controlled by the tip of the second arm 42a and presses the balloon 19. The number of second mechanisms 42 is the same as the number of balloons 19, and can be changed appropriately according to the number of balloons 19.

[0012] The culture device 5 adjusts the environment of the detachably held cell production cartridge 1. Specifically, for example, the culture device 5 adjusts the environment such as the temperature, humidity, and CO2 concentration of the culture section (not shown) housed in the hole 17H ​​of the held cell production cartridge 1.

[0013] FIG. 4 is a schematic diagram for explaining the function of the cell preparation cartridge of FIG. 1. The cell preparation cartridge 1 has a suspension supply unit 11, an inducer supply unit 12, a capture unit 13, an establishment unit 14, a washing liquid supply unit 15, a waste liquid storage unit 16, a culture unit 17, a medium supply unit 18, a balloon 19, a tube 20, a connector CN, and a valve VL as a closed flow path. The closed flow path has the suspension supply unit 11, the inducer supply unit 12, the establishment unit 14, the washing liquid supply unit 15, and the medium supply unit 18 as a storage unit that stores a fluid used in cell preparation and is at least partially movable. The diaphragm 3D transmits a driving force according to the energy to at least a part of the storage unit (the suspension supply unit 11, the inducer supply unit 12, the establishment unit 14, and the washing liquid supply unit 15). The balloon 19 transmits a driving force according to the energy to at least a part of the storage unit (the medium supply unit 18). The diaphragm 3D and the balloon 19 are movable parts, and transmit mechanical action according to the energy to the storage part via the movable parts. In addition, a part of the cell production cartridge 1 (the diaphragm 3D and the balloon 19) is structured to be deformable following the application of kinetic energy from the outside in order to transmit the energy for sending the fluid into the closed flow path. Similarly, at least a part of the storage part is structured to be deformable following the transmission of mechanical action from the energy transmission part. The suspension supply part 11, the inducer supply part 12, the capture part 13, the establishment part 14, the washing liquid supply part 15, the waste liquid storage part 16, the culture part 17, the medium supply part 18, the balloon 19, and the connector CN are housed in the holes 11H to 19H of the recess. The holes 11H to 19H may be formed with a support member (not shown) for supporting the contents such as a syringe or a bag. The tube 20 and the valve VL are housed in the groove 20T. The groove 20T may have a fitting portion into which the tube 20, the valve VL, the connector CN, etc. can be fitted. The tube 20 passes the fluid delivered from the reservoir. The connector CN detachably connects the tube 20 to the reservoir.

[0014] The closed flow path has, for example, a first flow path 21, a second flow path 22, and a third flow path 23 formed of a tube 20 that is opened or closed by pin members 31 to 38. The first flow path 21 is provided with a cleaning liquid supply unit 15, a suspension supply unit 11, a capture unit 13, an inducer supply unit 12, and a waste liquid storage unit 16 in the flow path direction (from upstream to downstream). The suspension supplied by the suspension supply unit 11 and the inducer supplied by the inducer supply unit 12 flow into the first flow path 21. In the first flow path 21, the suspension flows in a first direction, and the inducer flows in a second direction opposite to the first direction. The second flow path 22 is provided by branching off from the capture unit 13 of the first flow path 21 on the upstream side, and a base unit 14 is provided on the downstream side. The second flow path 22 is branched off from the first flow path 21 at a position between the suspension inlet and the capture unit 13 in the first flow path 21. The third flow path 23 is connected to the second flow path 22 on the downstream side, and has a medium supply section 18 provided on the upstream side and a culture section 17 provided on the downstream side.

[0015] The suspension supply unit 11 supplies a suspension containing target cells. The suspension may be blood in which target cells collected from a human body are suspended, or a preservation solution in which target cells are suspended. The target cells are cells that are to be initialized by an inducer. The target cells may be blood-derived cells or somatic cells derived from sources other than blood, so long as they are somatic cells that can be initialized. As the blood-derived target cells, mononuclear cells are used.

[0016] The suspension supply unit 11 is composed of an injector including a syringe and a piston, as shown in FIG. 5, for example. The syringe can contain the suspension and has a sealability capable of suppressing or preventing the suspension from being exposed to the outside air. The tip of the syringe is connected to the first flow path 21 via a connector CN and a tube 20 having a pin member 31 disposed thereon. The piston is pushed down by a rod-shaped member 41b via a diaphragm 3D to discharge the suspension contained in the syringe. The syringe is an example of a storage unit.

[0017] The inducer supplying section 12 supplies a medium containing an inducer to the capturing section 13. The medium containing an inducer includes an inducer and a medium for establishing induced pluripotent stem cells. The inducer initializes the target cells, and specifically, is an Oct family gene, a Klf family gene, a Myc family gene, or each of these gene products. As an example, Oct3 / 4 is used as the Oct family gene, Klf4 is used as the Klf family gene, and c-Myc or L-Myc is used as the Myc family gene. The inducer may also be a Sox family gene or a gene product thereof. As the Sox family gene, Sox2 is used.

[0018] The inducer supply unit 12 is composed of, for example, an injector including a syringe and a piston. The syringe can accommodate the inducer-containing medium and has a sealability capable of suppressing or preventing the inducer-containing medium from being exposed to the outside air. The tip of the syringe is connected to the first flow path 21 via a connector CN and a tube 20 having pin members 33 and 34 arranged above it. The piston is pushed down by the rod-shaped member 41b via the diaphragm 3D to discharge the inducer-containing medium accommodated in the syringe.

[0019] The capture unit 13 is provided in the first flow path 21 between the suspension supply unit 11 and the inducer supply unit 12. That is, the capture unit 13 is provided in the first flow path 21 between the inlet for the suspension and the inlet for the inducer. The capture unit 13 captures the target cells contained in the suspension supplied from the suspension supply unit 11. The capture unit 13 is composed of a container including a filter 131 that is capable of capturing the target cells and has openings that allow other substances having smaller particle sizes than the target cells to pass through.

[0020] The establishment section 14 is connected to a second flow path 22, and is supplied with target cells and induction factors via the second flow path 22, which includes a tube 20 with pin members 35, 36 arranged above. The establishment section 14 introduces the induction factors supplied to the capture section 13 into the target cells captured in the capture section 13, thereby establishing induced pluripotent stem cells (iPS cells). The establishment section 14 also moves the established induced pluripotent stem cells from the second flow path 22 to the culture section 17 via a third flow path 23, which includes a tube 20 with pin members 37 arranged above.

[0021] The establishment section 14 is composed of, for example, an injector including a syringe and a piston. The syringe can accommodate the induction factor-containing medium that is supplied from the induction factor supply section 12 and has passed through the capture section 13, and the target cells captured by the capture section 13, and has a sealability that can suppress or prevent the induction factor-containing medium and the target cells from being exposed to the outside air. The syringe can also accommodate iPS cells established by introducing an induction factor into the target cells. The tip of the syringe is connected to the second flow path 22 via the connector CN and the tube 20 with the pin member 36 disposed above. The piston is pushed down from the rod-shaped member 41b via the diaphragm 3D to discharge the iPS cells accommodated in the syringe.

[0022] The washing liquid supply unit 15 supplies a washing liquid to the first flow path 21, and the washing liquid washes the capture unit 13. The washing liquid is, for example, physiological saline, etc., and a liquid that causes little damage to the target cells is used.

[0023] The cleaning liquid supply unit 15 is composed of, for example, an injector including a syringe and a piston. The syringe can contain the cleaning liquid and has a sealability capable of suppressing or preventing the cleaning liquid from being exposed to the outside air. The tip of the syringe is connected to the first flow path 21 via a connector CN and a tube 20 having a pin member 32 disposed above it. The piston is pushed down by the rod-shaped member 41b via the diaphragm 3D to discharge the cleaning liquid contained in the syringe.

[0024] The waste liquid storage section 16 is a container that stores waste liquid. The waste liquid storage section 16 stores the suspension and washing liquid that have passed through the capture section 13 as waste liquid. The waste liquid storage section 16 may be a container that can store waste liquid, such as a flask or a bag.

[0025] The culture section 17 cultures the iPS cells established by the establishment section 14. The culture section 17 may be a container capable of culturing iPS cells, such as a well plate, a flask, a dish, or a bag.

[0026] The medium supplying unit 18 supplies medium for culturing iPS cells to the culture unit 17 via a third flow path 23 connected via a tube 20 with a pin member 38 disposed above. The medium supplying unit 18 is a container for storing the medium, and a container such as a bag can be used.

[0027] As shown in FIG. 6, the culture medium supply unit 18 is composed of a bag having tubes at the upper and lower ends. The bag can contain the culture medium and has a sealability that can suppress or prevent the culture medium from being exposed to the outside air. The tube at the lower end of the bag is connected to the third flow path 23 via a connector CN and a tube 20 with a pin member 38 disposed thereon. The tube at the upper end of the bag is connected to the balloon 19 via a connector CN and a valve VL. When a detachable handle 39 is attached, the valve VL can be opened and closed in response to the operation of the handle 39. The balloon 19 is pressed by the columnar member 42b, and introduces the contained air to the upper end of the culture medium supply unit 18 via the valve VL, the connector CN, and the tube. The culture medium supply unit 18 supplies the culture medium pressed by the introduced air from the tube at the lower end to the third flow path 23.

[0028] Moreover, pin members 31 to 34 are provided above the elastically deformable tube 20 constituting the first flow path 21 so as to be movable in the vertical direction. The pin members 31 and 32 are disposed above the tube 20 on one end side (upstream side) of the first flow path 21, and the suspension supply unit 11 and the cleaning liquid supply unit 15 are connected to the tube 20. The pin member 31 can switch between supplying and stopping the suspension supplied from the suspension supply unit 11 to the first flow path 21 by opening the tube 20 as shown in FIG. 7 or closing the tube 20 as shown in FIG. 8. Note that FIG. 7(a) shows a cross section substantially parallel to the tube 20, and FIG. 7(b) shows a cross section substantially perpendicular to the tube 20, and the tip 31a of each pin member 31 is not in contact with the tube 20. The pin member 31 may be, for example, a ball pin having a ball 31b biased outward. The ball 31b functions as a positioning member by engaging with a hole 20Ta formed in the groove 20T. Similarly, FIG. 8(a) shows a cross section approximately parallel to the tube 20, and FIG. 8(b) shows a cross section approximately perpendicular to the tube 20, in which the tip 31a of the pin member 31 crushes a part of the tube 20 to block the flow path. The hole 20Ta formed in the groove 20T is formed corresponding to the position where the pin member 31 blocks the tube 20. The other pin members 32 to 38 function in the same manner as the pin member 31 shown in FIG. 7 and FIG. 8. The pin member 32 can switch between supplying and stopping the cleaning liquid supplied from the cleaning liquid supply unit 15 to the first flow path 21 by opening or closing the tube 20. The pin member 33 is disposed above the tube 20 on the other end side (downstream side) of the first flow path 21, and the inducer supply unit 12 is connected to the tube 20. The pin member 33 can switch between supplying and stopping the medium containing the inducer to the first flow path 21 by opening or closing the tube 20. In addition, in the first flow path 21, a pin member 34 is disposed above the tube 20 connected to the inlet side of the waste liquid storage section 16. The pin member 34 makes it possible to switch between supplying and stopping the suspension and washing liquid that have passed through the capture section 13 to the waste liquid storage section 16 by opening or closing the tube 20.

[0029] Pin members 35 to 37 are provided above the tube 20 constituting the second flow path 22. The pin member 35 can switch between supplying and stopping the medium containing the induction factor that has passed through the capture section 13 and the target cells captured by the capture section 13 to the second flow path 22 by opening or closing the tube 20. The pin members 36 and 37 can switch between supplying and stopping the induced pluripotent stem cells established in the establishment section 14 to the third flow path 23 by opening or closing the tube 20. The pin member 38 can switch between supplying and stopping the medium supplied from the medium supply section 18 to the third flow path 23 by opening or closing the tube 20. Such pin members 31 to 38 are held by the container body 2 and the lid 3 so as to be movable along a direction perpendicular to the flow path, and are an example of a blocking member that blocks the flow path by elastically deforming a part of the flow path.

[0030] The tube 20 constituting the first flow path 21, the second flow path 22, and the third flow path 23 is made of a tubular member having sealing properties and flexibility, such as a vinyl or plastic tube, etc. This suppresses or prevents various liquids passing through the first flow path 21, the second flow path 22, and the third flow path 23 from being exposed to the outside air.

[0031] An example of the operation of the cell production cartridge and its peripheral components configured as above will be described with reference to the flow chart in FIG. 9 and the schematic diagrams in FIG. 10 to FIG. 14. In the following description, it is assumed that the suspension is blood collected from a donor. In the cell production cartridge 1, it is assumed that the suspension supply unit 11 stores one dose of blood, the inducer supply unit 12 stores one dose of culture medium containing an inducer, and the cleaning solution supply unit 15 stores one dose of cleaning solution. That is, the cell production cartridge 1 is disposable and is used only for one cell production process. In addition, in the cell production cartridge 1, it is assumed that the pin members 31 to 38 block the flow paths 21 to 23, and the valve VL is in a closed state. In addition, each step ST11 to ST16 in the cell production process is performed by an automatic mechanism including the liquid sending mechanism 4, the culture device 5, the transport device, the switching device, and the control device without the intervention of a user. However, each step ST11 to ST16 may be performed by the intervention of a user as necessary.

[0032] In step ST11, the cell production cartridge 1 is transported below the liquid sending mechanism 4 by a transport device (not shown) and set (attached) to the culture device 5. Thereafter, a switching device (not shown) capable of switching the pin members 31 to 38 and the handle 39 is attached to the cell production cartridge 1.

[0033] After step ST11, in step ST12, the insertion positions of the pin members 31-38 of the cell production cartridge 1 are controlled by a switching device (not shown) to switch between an open state and a closed state of the tube 20. As a result, the flow paths 21-23 are switched.

[0034] After step ST12, in step ST13, the liquid sending mechanism 4 applies energy to the cell production cartridge 1. For example, the first mechanism 41 presses the piston via the diaphragm 3D. Alternatively, the second mechanism 42 presses the balloon 19.

[0035] After step ST13, in step ST14, the cell production cartridge 1 sends out the liquid along the switched flow path in response to the applied energy.

[0036] After step ST14, in step ST15, a control device (not shown) judges whether or not the cell production process has been completed. If the result of this judgment is no, the control device returns to step ST12 and controls the switching device and the liquid sending mechanism 4 to repeat the processes of steps ST12 to ST15. On the other hand, if the result of the judgment in step ST14 is that the cell production process has been completed, the control device starts up the culture device 5 and proceeds to step ST16.

[0037] After step ST15, in step ST16, the culture device 5 adjusts the environment of the culture section 17 housed in the cell production cartridge 1. This cultures the cells in the culture section 17. If necessary, the control device also supplies culture medium from the culture medium supply section 18 of the cell production cartridge 1 to the culture section 17 by controlling the switching device and the liquid sending mechanism 4. Thereafter, when the culture is completed, the cell production process is completed.

[0038] Next, among the above operations, an example of the repeating operations of steps ST12 to ST14 related to the flow path of the cell production cartridge 1 and the operation of step ST16 will be described.

[0039] (First steps ST12~ST14) As shown in Fig. 10, the cell production cartridge 1 is raised by a switching device (not shown) so that the tip positions of the pin members 31 and 34 open the tube 20. As a result, the flow path of the cell production cartridge 1 is switched so that the suspension supply unit 11 communicates with the waste liquid storage unit 16 via the capture unit 13 and the flow path 21 (step ST12). In Fig. 10, in the circles representing the pin members 31 to 38, a straight line overlapping the flow path represents an open state of the flow path, and a straight line perpendicular to the flow path represents a closed state of the flow path. The same applies to the other figures.

[0040] The liquid sending mechanism 4 applies energy to the suspension supplying part 11 by the first mechanism 41 pressing the piston of the suspension supplying part 11 via the diaphragm 3D (step ST13).

[0041] The suspension supply unit 11 sends out the suspension along the switched flow path in response to the applied energy (step ST14). At this time, the suspension supply unit 11 supplies the suspension (blood) containing the target cells to the capture unit 13 in the forward direction via the first flow path 21. The forward direction is defined as the direction in which the suspension moves toward the waste liquid storage unit 16. Specifically, the suspension supply unit 11 discharges one batch of the suspension containing the target cells required for producing iPS cells into the first flow path 21. The suspension discharged into the first flow path 21 flows into the capture unit 13. Among various components (substances) contained in the suspension flowing into the capture unit 13, components having a particle size larger than the opening of the filter 131 are captured, and components having a particle size smaller than the opening of the filter 131 pass through the filter 131. As the filter 131, for example, a filter having an opening capable of capturing components of the same size as white blood cells is preferably used. The target cells have a particle size larger than the opening, and are captured by the filter 131. Red blood cells and the like that are not target cells have a particle size smaller than the openings and therefore pass through filter 131. The blood components that have passed through capture section 13 are stored in waste liquid storage section 16 as waste liquid.

[0042] (Second steps ST12~ST14) 11, in the cell production cartridge 1, a switching device (not shown) pushes down the tip position of the pin member 31 so as to close the tube 20, and then the tip position of the pin member 32 is pulled up so as to open the tube 20. As a result, the flow path of the cell production cartridge 1 is switched so that the washing liquid supply unit 15 communicates with the waste liquid storage unit 16 via the capture unit 13 and the flow path 21 (step ST12).

[0043] The liquid sending mechanism 4 applies energy to the cleaning liquid supply part 15 by the first mechanism 41 pressing the piston of the cleaning liquid supply part 15 via the diaphragm 3D (step ST13).

[0044] The washing liquid supply unit 15 sends out the washing liquid along the switched flow path in response to the applied energy (step ST14). At this time, the washing liquid supply unit 15 flushes the unnecessary blood components other than the target cells remaining in the first flow path 21 and the capture unit 13 into the waste liquid storage unit 16 with the washing liquid. The unnecessary blood components are mainly plasma components, red blood cell components, and platelet components. The unnecessary blood components entangled in the first flow path 21 and the capture unit 13 may be separated and efficiently washed away by repeatedly pushing and pulling the piston of the washing liquid supply unit 15 to eject and suck up the washing liquid.

[0045] (Third steps ST12~ST14) 12, in the cell production cartridge 1, a switching device (not shown) pushes down the tip positions of the pin members 32 and 34 so as to close the tube 20, and then the tip positions of the pin members 33, 35 and 36 are pulled up so as to open the tube 20. As a result, the flow path of the cell production cartridge 1 is switched so that the inducer supplying section 12 communicates with the establishment section 14 via the capture section 13 and the flow paths 21 and 22 (step ST12).

[0046] In the liquid sending mechanism 4, the first mechanism 41 presses the piston of the inducer supplier 12 via the diaphragm 3D, thereby applying energy to the inducer supplier 12 (step ST13).

[0047] The inducer supplying unit 12 delivers the inducer-containing medium along the switched flow path in response to the applied energy (step ST14). At this time, the inducer supplying unit 12 supplies the inducer-containing medium to the capture unit 13 via the first flow path 21 in the direction opposite to the forward direction, and supplies the inducer-containing medium and the target cells to the establishment unit 14 via the second flow path 22. Specifically, the inducer supplying unit 12 discharges the stored inducer-containing medium for one use. The discharged inducer-containing medium flows into the capture unit 13 in the opposite direction to the flow direction of the suspension. The inducer-containing medium that flows into the capture unit 13 passes through the filter 131 and flows into the establishment unit 14 via the second flow path 22 together with the target cells captured by the filter 131. In order to accommodate the inducer-containing medium and the target cells in the syringe of the establishment unit 14, it is advisable to retract the piston of the establishment unit 14 to its limit in advance.

[0048] In the establishment section 14, an inducer is introduced into the target cells, and an iPS cell is established from the target cells. The inducer can be supplied in various forms. For example, the inducer may be incorporated into various vectors and supplied. The vector is not particularly limited, and may be a viral vector such as a Sendai virus vector or a retrovirus vector, or a non-viral vector such as a plasmid. For example, when a Sendai virus vector incorporating an inducer is used, the inducer is introduced into the target cells by contacting the Sendai virus vector with the target cells. Thereafter, a gene product of the inducer is synthesized. The action of the gene product induces initialization of the target cells, and iPS cells, which are undifferentiated cells having pluripotency and proliferation ability, are generated. The iPS cells are cultured for about 1 hour in the presence of a culture medium. This allows the establishment of iPS cells. During the establishment operation, the establishment section 14 may be kept at about 37 degrees and 5% CO2.

[0049] In order to improve the accuracy of establishment of iPS cells in the establishment section 14, it is advisable to use a container with a relatively small volume as the syringe of the establishment section 14. This increases the probability of contact between the target cells and the inducer or the vector incorporating the inducer, and thus increases the probability of establishing iPS cells.

[0050] (4th step ST12~ST14) 13, in the cell production cartridge 1, a switching device (not shown) pushes down the tip positions of the pin members 33 and 35 so as to close the tube 20, and then the tip position of the pin member 37 is pulled up so as to open the tube 20. As a result, the flow paths of the cell production cartridge 1 are switched so that the establishment section 14 communicates with the culture section 17 via the flow paths 22 and 23 (step ST12).

[0051] In the liquid delivery mechanism 4, the first mechanism 41 presses the piston of the establishment portion 14 via the diaphragm 3D, thereby applying energy to the establishment portion 14 (step ST13).

[0052] The establishment section 14 sends out the established iPS cells along the switched flow path in response to the applied energy (step ST14). At this time, the establishment section 14 discharges the iPS cell suspension into the second flow path 22. The iPS cell suspension discharged into the second flow path 22 moves to the culture section 17.

[0053] (Step ST16) The culture device 5 adjusts the environment of the culture section 17 housed in the cell production cartridge 1 to room temperature of about 37 degrees. The culture section 17 cultures the transferred iPS cells. If necessary, the control device controls the switching device and the liquid sending mechanism 4 to supply the culture medium from the culture medium supply section 18 of the cell production cartridge 1 to the culture section 17. For example, as shown in FIG. 14, the cell production cartridge 1 is pushed down by a switching device (not shown) so that the tip positions of the pin members 36 and 37 close the tube 20, and then the tip position of the pin member 38 is raised so that the tube 20 is opened. As a result, the flow path of the cell production cartridge 1 is switched so that the culture medium supply section 18 communicates with the culture section 17 via the flow path 23. Also, for example, the cell production cartridge 1 is operated by a switching device (not shown) so that the handle 39 opens the valve VL. As a result, the flow path of the cell production cartridge 1 is switched so that the balloon 19 communicates with the culture medium supply section 18 via the valve VL.

[0054] In the liquid delivery mechanism 4, the second mechanism 42 presses the balloon 19, thereby applying energy to the culture medium supply unit 18.

[0055] The culture medium supplying unit 18 sends out the culture medium along the switched flow path in response to the applied energy. At this time, the culture medium supplying unit 18 discharges the culture medium into the third flow path 23. The culture medium discharged into the third flow path 23 moves to the culture unit 17. Thereafter, when the culture is finished, the cell production process is finished.

[0056] As described above, according to one embodiment, the cell preparation cartridge 1 includes a flow path and a container that houses the flow path. The flow path may be a closed system flow path. The container may be a closed system container. The container includes an energy transmission unit that receives energy from the outside of the container for sending a fluid in the flow path. The energy transmission unit is located at a boundary that separates the inside and outside of the container. Therefore, even if the closed system flow path is damaged and the fluid leaks, the closed system container does not allow the leaked fluid to flow out. Therefore, safety can be ensured without making capital investments associated with the possibility of liquid leaking. To supplement this, according to one embodiment, safety can be ensured locally within the cell preparation cartridge 1, not in global facilities such as a safety cabinet or a laboratory. Also, according to one embodiment, if the cell preparation cartridge 1 is positioned and placed in the culture device 5, the contained closed system flow path and the energy transmission unit for sending the fluid are also positioned and placed collectively, so that operability can be improved. That is, according to one embodiment, a double closed structure such as a closed system flow path and a closed system container that houses the flow path can achieve both safety and operability. In addition, since the operability can be improved and the associated capital investment can be omitted, high-quality and inexpensive cell production becomes possible. Furthermore, when the cell production cartridge 1 is set in an automatic mechanism and processes that require the skill of the user, such as the process of sending fluid, the process of switching flow paths, and the process of culturing cells, are automated, stable quality can be ensured regardless of the skill of the user.

[0057] According to one embodiment, the flow path has a storage section that stores a fluid used in cell production and has at least a portion that is movable. The energy transmission section transmits a driving force corresponding to the energy to at least a portion of the storage section. Therefore, in addition to the above-mentioned effects, when the storage section receives the driving force corresponding to the energy, the storage section operates according to the driving force to eject the fluid, so that the fluid can be pumped within the fluid.

[0058] According to one embodiment, the energy transmission unit is a movable part, and transmits a mechanical action corresponding to the energy to the storage unit via the movable part. Therefore, in addition to the above-mentioned effects, the configuration in which the movable part transmits a mechanical action corresponding to the energy reduces the loss when converting the energy into a mechanical action, and therefore it is possible to save the energy applied to the cell production cartridge 1.

[0059] According to one embodiment, the reservoir is a syringe, and therefore in addition to the above-mentioned effects, precise quantitative liquid delivery can be achieved.

[0060] According to one embodiment, the energy transmission unit is a diaphragm 3D. Therefore, in addition to the above-mentioned effects, when the amount of liquid to be delivered is relatively small, the energy transmission unit can be realized with a simple configuration of a diaphragm with a small amount of displacement.

[0061] Moreover, according to one embodiment, the energy transmission part is the balloon 19. Therefore, in addition to the above-mentioned effects, it is possible to realize simple liquid delivery without the need for a fixed amount.

[0062] According to one embodiment, the container in the cell production cartridge 1 includes a container body 2 having a recess formed therein for holding a closed flow path, and a lid 3 attached to the container body 2 so as to cover the recess. The lid 3 includes an energy transmission unit at a position covering a storage unit in the flow path. Therefore, in addition to the above-mentioned effect, the storage unit and the energy transmission unit can be arranged to face each other simply by closing the lid 3 of the container body 2, and therefore the positioning of the storage unit and the energy transmission unit can be easily performed.

[0063] Furthermore, according to one embodiment, the pin members 31-38 are held by the container body 2 and the lid 3 so as to be movable along a direction perpendicular to the flow path, and close the flow path by elastically deforming a part of the flow path. Therefore, in addition to the above-mentioned effects, the flow path can be closed with a simple configuration in which the pin members 31-38 crush the tube 20.

[0064] According to one embodiment, the flow path includes a tube through which the fluid delivered from the reservoir passes, and a connector that detachably connects the tube to the reservoir, thereby providing the above-mentioned effects, and allowing any reservoir to be detached from the flow path via the connector.

[0065] (Variation 1) In one embodiment, the cell production cartridge 1 includes a diaphragm 3D to receive energy from the liquid delivery mechanism 4, but is not limited thereto. For example, the cell production cartridge 1 may include a bellows 3B instead of the diaphragm 3D as shown in FIG. 15. Even with such a modified example, in addition to the same effects as in the one embodiment, when the amount of liquid delivered is relatively large, an energy transmission section can be realized with a simple configuration of a bellows with a large amount of displacement. The bellows 3B is another example of an energy transmission section.

[0066] (Variation 2) In one embodiment, a finished cell preparation cartridge 1 manufactured by a manufacturer or the like is used, but the present invention is not limited thereto. For example, as shown in FIG. 16, the cell preparation cartridge 1 may be assembled by a user. In FIG. 16, the user prepares a container body 2 that houses a closed flow path to which downstream parts such as an establishment part 14, a waste liquid storage part 16, and a culture part 17 are connected (step ST1). In step ST1, the user may house a closed flow path in the container body 2 manufactured by a manufacturer or the like. Alternatively, the user may obtain a container body 2 that houses a closed flow path manufactured by a manufacturer or the like. Next, the user connects the inducer supply part 12, the cleaning liquid supply part 15, and the medium supply part 18 to the connector CN of the flow path (step ST2). In addition, the user connects the balloon 19 filled with air to the upper connector CN of the medium supply part 18 with the valve VL closed (step ST3). In addition, the user connects the suspension supply part 11 to the connector CN of the flow path (step ST4). As a result, a closed flow path is formed in the container body 2. Next, the user closes the lid 3 of the container body 2. As a result, a closed container containing a closed flow path is formed (step ST5). Thereafter, the user attaches the pin members 31-38 and the handle 39 to the closed container (step ST6). This completes the assembly of the cell production cartridge 1 (step ST7). In this modified example, in addition to the effects of the embodiment, it is possible to use the suspension supply unit 11, the washing liquid supply unit 15, the inducer supply unit 12, and the culture medium supply unit 18 that store the liquid that the user wants to use.

[0067] (Variation 3) In one embodiment, the suspension supply unit 11, the inducer supply unit 12, the establishment unit 14, and the cleaning solution supply unit 15 are configured with a syringe and a piston, but are not limited to this. For example, the suspension supply unit 11, the inducer supply unit 12, the establishment unit 14, and the cleaning solution supply unit 15 may be flexible containers to which balloons 19 are appropriately connected. Flexible plastic or vinyl bags are suitable as flexible containers. Even with such a modified example, the same effects as those of the first embodiment can be obtained.

[0068] (Variation 4) In one embodiment, the culture medium supply unit 18 is configured with a plastic or vinyl bag and is connected to the balloon 19, but is not limited to this. For example, the culture medium supply unit 18 may be configured with a syringe and a piston, and may be unrelated to the balloon 19. Even with such a modified example, the same action and effect as in the first embodiment can be obtained.

[0069] (Variation 5) In the embodiment, the flow path is closed by crushing a part of the tube 20 with the pin members 31 to 38, but the present invention is not limited to this. For example, a solenoid valve may be connected to the tube 20, and the flow path may be opened or closed according to the control of the solenoid valve. Even with such a modified example, the same effect as that of the embodiment can be obtained.

[0070] (Variation 6) In the embodiment, the flow path is switched by any of the pin members 31 to 38 appropriately pressing the tube 20, but the present invention is not limited to this. For example, instead of the pin members 31 to 38 arranged above the tube 20, a three-way valve may be arranged at a branching portion of the tube 20, and the flow path may be switched by the three-way valve. Even with such a modified example, the same action and effect as the embodiment can be obtained.

[0071] (Variation 7) In one embodiment, the cell production cartridge 1 receives kinetic energy from the liquid delivery mechanism 4 as energy for delivering the fluid in the flow path, but is not limited to this. For example, the cell production cartridge 1 may receive any one of potential energy, thermal energy, electrical energy, light energy, and chemical energy from the liquid delivery mechanism 4 as energy for delivering the fluid in the flow path. The cell production cartridge 1 is provided with a mechanism for generating a pressure gradient in the flow path from the received energy, so that it can deliver the fluid according to the received energy.

[0072] (Variation 8) In one embodiment, the cell preparation cartridge 1 has a horizontally elongated rectangular parallelepiped shape in which the top and bottom faces of the six rectangular faces constituting the rectangular parallelepiped are larger than the other four faces, but is not limited thereto. For example, the cell preparation cartridge 1 may have a vertically elongated rectangular parallelepiped shape in which the two opposing side faces of the six rectangular faces constituting the rectangular parallelepiped are larger than the other four faces. Even with such a modified example, the same action and effect as the one embodiment can be obtained.

[0073] (Variation 9) In one embodiment, the cell preparation cartridge 1 has a layout capable of accommodating only one type of closed-system flow path, but is not limited thereto. For example, the cell preparation cartridge 1 may have a layout capable of accommodating two types of closed-system flow paths. In this case, the container body 2 of the cell preparation cartridge 1 has a layout consisting of a recess used for both of the two flow paths, a recess used only for one of the flow paths, and a recess used only for the other flow path. Similarly, the cell preparation cartridge 1 may have a layout capable of accommodating three or more types of closed-system flow paths. According to such a modified example, in addition to the effect of one embodiment, the versatility of the cell preparation cartridge 1 can be improved.

[0074] (Variation 10) In one embodiment, the cell production cartridge 1 has a double closed structure including a closed flow path and a closed container that houses the flow path, but is not limited thereto. For example, the cell production cartridge 1 may have a triple closed structure including a container that further houses the double closed structure. Alternatively, the cell production cartridge 1 is not limited to a triple closed structure, and may have a quadruple or more multiple closed structure. According to such a modified example, in addition to the effect of the one embodiment, it is expected that the closed flow path can be protected from impact when the cell production cartridge 1 is dropped, and therefore safety can be further improved.

[0075] (Variation 11) In one embodiment, the entire surface of the lid 3 is transparent, but this is not limiting. For example, the entire surface of the lid 3 may be opaque, or only a specific area may be transparent. Also, a light-shielding filter may be removably attached to the lid 3, so that the transparent area can be shielded from light as necessary. This makes it possible to store liquid that requires light shielding, or to culture cells in a light-shielding environment.

[0076] (Variation 12) In one embodiment, the entire surface of the container body 2 is opaque, but this is not limited thereto. For example, the entire surface of the container body 2 may be transparent, or only a specific region may be transparent. In addition, when the entire surface of the container body 2 is transparent, a light-shielding filter may be detachably attached only to a specific surface of the container body 2 or only to a partial region of the specific surface, so that the transparent region can be shielded from light as necessary. This makes it possible to store a liquid that requires light shielding, or to culture cells in a light-shielding environment.

[0077] According to at least one of the embodiments described above, safety can be ensured without making capital investments associated with the possibility of liquid leakage.

[0078] Although some embodiments have been described, these embodiments are presented as examples and are not intended to limit the scope of the invention. These embodiments can be implemented in various other forms, and various omissions, substitutions, modifications, and combinations of embodiments can be made without departing from the spirit of the invention. These embodiments and their modifications are included in the scope of the invention and its equivalents as described in the claims, as well as in the scope and spirit of the invention.

[0079] Regarding the above embodiment, the following supplementary notes are disclosed as one aspect and optional features of the invention. (Supplementary Note 1) A cartridge for cell preparation comprising a flow path and a container for accommodating the flow path.

[0080] The container may include an energy transmission section that receives energy for transporting fluid through the flow path from an outside of the container.

[0081] The energy transfer portion may be located at a boundary separating the inside and outside of the container. (Appendix 2) The flow channel may have a reservoir portion that stores a fluid used in cell production and at least a portion of which is movable.

[0082] The energy transmission unit may transmit a driving force corresponding to the energy to at least the part of the storage unit. (Appendix 3) The energy transmission portion may be a movable part, and may transmit a mechanical action corresponding to the energy to the storage portion via the movable part. (Appendix 4) The reservoir may be a syringe. (Appendix 5) The energy transfer portion may be a diaphragm. (Appendix 6) The energy transfer portion may be a bellows. (Appendix 7) The energy delivery portion may be a balloon. (Appendix 8) The container comprises: A container body having a recess for holding the flow path; a lid attached to the container body so as to cover the recess; The present invention may also include:

[0083] The lid may include the energy transfer portion at a position of the flow path that covers the storage portion. (Appendix 9) The liquid supply device may further include a blocking member that is held by the container body and the lid so as to be movable along a direction perpendicular to the flow path, and that blocks the flow path by elastically deforming a portion of the flow path. (Appendix 10) The flow path may further include a tube through which the fluid delivered from the reservoir passes, and a connector that detachably connects the tube to the reservoir. (Appendix 11) The flow path may be a closed flow path. (Appendix 12) The container may be a closed container. [Explanation of symbols]

[0084] 1. Cell Preparation Cartridge 2 Container body 3 Lid 3a, 11H~19H, 20Ta hole 3B Bellows 3D Diaphragm 4. Liquid delivery mechanism 5 Culture device 11 Suspension supply section 12 Inducer supply section 13 Capture unit 131 Filters 14 Establishment 15 Cleaning liquid supply unit 16 Waste liquid storage section 17 Culture Department 18 Culture medium supply section 19 Balloon 20 Tubes 20T groove 21 First Channel 22 Second Channel 23 The third channel 31~38 Pin parts 39 Handle 41 Mechanism 1 41a 1st Arm 41b Rod-shaped member 42 Second mechanism 42a 2nd Arm 42b Column member

Claims

1. A cell preparation cartridge including a flow path and a container for accommodating the flow path, the container includes an energy transfer unit that receives energy for transferring a fluid through the flow path from an outside of the container; The energy transfer unit is located at the boundary separating the inside and the outside of the container. Cartridge for cell preparation.

2. The flow channel has a reservoir portion that stores a fluid used in cell production and is at least partially movable; The energy transmission unit transmits a driving force corresponding to the energy to at least a portion of the storage unit. The cell preparation cartridge according to claim 1 .

3. The energy transfer unit is a movable part, and transfers a mechanical action corresponding to the energy to the storage unit via the movable part. The cell preparation cartridge according to claim 2 .

4. The cell production cartridge according to claim 2 , wherein the storage portion is a syringe.

5. The cell production cartridge according to claim 2 , wherein the energy transmission part is a diaphragm.

6. The cell production cartridge according to claim 2 , wherein the energy transmission part is a bellows.

7. The cell production cartridge according to claim 2 or 3, wherein the energy transmission part is a balloon.

8. The container comprises: A container body having a recess for holding the flow path; a lid attached to the container body so as to cover the recess; It is equipped with The cell production cartridge according to claim 2 , wherein the lid is provided with the energy transmission section at a position covering the reservoir section of the flow channel.

9. The cell production cartridge of claim 8, further comprising a blocking member that is held by the container body and the lid so as to be movable along a direction perpendicular to the flow path, and that blocks the flow path by elastically deforming a portion of the flow path.

10. The cell production cartridge according to any one of claims 2 to 4, wherein the flow path further comprises a tube through which fluid discharged from the storage portion passes, and a connector for detachably connecting the tube to the storage portion.

11. The cell production cartridge according to claim 1 , wherein the flow path is a closed system flow path.

12. The cell preparation cartridge according to claim 1 or 11, wherein the container is a closed container.

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