Perfusion culture device and perfusion culture system

Abstract

To provide a perfusion culture apparatus and a perfusion culture system which can suppress mixing of foreign matters into a culture medium while simplifying the apparatus configuration.SOLUTION: A perfusion culture apparatus 1 comprises: an outer vessel 5 which has six wells 5a housing a culture medium; six culture vessels 8 which are cylindrical and whose end parts on a bottom side of the wells 5a in the state of being inserted into the wells 5a are obstructed by a porous film 802 disseminated with a cell or a biological tissue; a vessel plate 7 which is arranged facing the outer vessel 5, and in which holding holes 7a, which hold the culture vessels 8 by their inner sides in portions facing each of the six wells 5a, are formed; and a drive part 11 which moves the vessel plate 7 with respect to the outer vessel 5 such that it alternately moves in a direction where the culture vessels 8 held by the vessel plate 7 approach the bottom of the wells 5a and in a direction where the culture vessels 8 go away from the bottom of the wells 5a.SELECTED DRAWING: Figure 1

Description

Technical Field

[0004] , , , ,

[0001] The present invention relates to a perfusion culture device and a perfusion culture system.

Background Art

[0002] An automatic perfusion culture device has been proposed in which a culture vessel having a porous membrane at the bottom, into which cells are seeded, in each well of a well plate containing a medium is inserted, and the medium is perfused in each well and the culture vessel to perform cell culture (see, for example, Patent Document 1). This automatic perfusion culture device includes a supply tube provided with a supply nozzle at its tip and arranged to supply the medium into the well, a supply pump connected to the side opposite to the supply nozzle side of the supply tube and supplying the medium into the well through the supply tube and the supply nozzle, a discharge tube provided with a discharge nozzle at its tip and arranged to discharge the medium from the culture vessel, and a discharge pump discharging the medium from the culture vessel through the discharge nozzle and the discharge tube.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0004] However, in the automated perfusion culture apparatus described in Patent Document 1, it is necessary to provide one set of supply tubes and discharge tubes for each well. Therefore, as the number of wells in the well plate increases, the number of supply tubes and discharge tubes must also increase accordingly, complicating the apparatus structure. Furthermore, in this automated perfusion culture apparatus, the supply pump supplies the culture medium from the culture medium supply container to the wells through multiple supply tubes. Therefore, connectors must be interposed at least between the culture medium supply container and the supply pump, and between the supply pump and the supply tubes. As a result, there is a risk that microorganisms or other foreign matter may enter the supply pump or supply tubes from the outside through the connector.

[0005] This invention has been made in view of the above reasons, and aims to provide a perfusion culture apparatus and perfusion culture system that can simplify the apparatus configuration while suppressing the contamination of the culture medium with foreign matter. [Means for solving the problem]

[0006] To achieve the above objective, the perfusion culture apparatus according to the present invention is An outer container having multiple wells for containing culture medium, Multiple culture vessels, which are cylindrical in shape and whose bottom end, when inserted into the wells, is closed with a porous membrane into which cells or biological tissues are seeded, A container plate that is plate-shaped and arranged opposite to the side in which each of the multiple wells of the outer container opens, and having holding holes for holding the culture container on the inside provided in the portion opposite to each of the multiple wells, The system includes a drive unit that moves at least one of the container plate and the outer container relative to the other such that at least one of the culture containers held on the container plate moves alternately in a first direction toward the bottom of the well and in a second direction toward away from the bottom of the well. [Effects of the Invention]

[0007] According to the present invention, the container plate has holding holes formed on the inside of each of the multiple wells, and the drive unit moves at least one of the container plate and the outer container relative to the other such that at least one culture container held by the container plate moves alternately in a first direction toward the bottom of the well and in a second direction toward away from the bottom of the well. As a result, by simply moving the container plate relative to the outer container, a pressure difference can be generated between the inside and outside of the culture containers in their porous membranes in multiple culture containers, and the culture medium can be perfused in each of the multiple culture containers simultaneously. Thus, tubes and pumps for perfusing the culture medium in the wells of the outer container and in the culture containers are not required, and there is an advantage that cell or biological tissue perfusion culture can be performed with a simple configuration consisting of a container plate and a drive unit that drives the container plate. Furthermore, since it is not necessary to continuously supply the culture medium from the outside during cell or biological tissue perfusion culture, contamination of the culture medium with microorganisms and other foreign substances when supplying the culture medium from the outside can be suppressed. [Brief explanation of the drawing]

[0008] [Figure 1] The perfusion culture apparatus according to Embodiment 1 of the present invention is shown, where (A) is a plan view, (B) is a cross-sectional view along line AA of (A), and (C) is a cross-sectional view along line BB of (A). [Figure 2] This is a cross-sectional view of the perfusion culture apparatus according to Embodiment 1, where (A) shows the container plate moving away from the outer container, and (B) shows the container plate moving towards the outer container. [Figure 3] This is an explanatory diagram of the operation of a perfusion culture vessel according to Embodiment 1, where (A) shows the time profile of the displacement of the container plate relative to the outer container, (B) shows the time profile when the amplitude of the displacement is halved and the fluctuation period is halved, and (C) shows the time profile when the fluctuation period is halved. [Figure 4] This is a cross-sectional view of the perfusion culture system according to Embodiment 2. [Figure 5] This is a plan view of the perfusion culture system according to Embodiment 3. [Figure 6] This is a cross-sectional view of the perfusion culture system according to Embodiment 3. [Figure 7] This flowchart shows an example of the flow of cell culture processing performed by the perfusion culture system according to Embodiment 3. [Figure 8] The perfusion culture system according to Embodiment 3 is shown, where (A) is a cross-sectional view and (B) is a plan view. [Figure 9] The perfusion culture system according to Embodiment 3 is shown, where (A) is a plan view and (B) is a cross-sectional view. [Figure 10] The perfusion culture system according to Embodiment 3 is shown, where (A) is a plan view and (B) is a cross-sectional view. [Figure 11] This flowchart shows an example of the flow of cell culture processing performed by the perfusion culture system according to Embodiment 3. [Figure 12] The perfusion culture system according to Embodiment 3 is shown, where (A) is a plan view and (B) is a plan view in another state. [Figure 13] This is a cross-sectional view of a modified perfusion culture apparatus. [Figure 14] This is a cross-sectional view of a modified perfusion culture apparatus. [Modes for carrying out the invention]

[0009] (Embodiment 1) The separation member according to an embodiment of the present invention will now be described with reference to the drawings. The perfusion culture apparatus according to this embodiment comprises an outer container having a plurality of wells for containing a culture medium, a plurality of culture vessels which are cylindrical and whose ends on the bottom side of the wells when inserted into the wells are closed with a porous membrane on which cells or biological tissues are seeded, a container plate, and a drive unit for driving the container plate. The container plate is plate-shaped and is positioned opposite each of the plurality of wells in the outer container to the side where each well opens, and has holding holes that hold the culture vessels on the inside through the portion opposite each of the plurality of wells. The drive unit moves at least one of the container plate and the outer container relative to the other so that at least one culture vessel held by the container plate moves alternately in a first direction in which it is inserted into the inside of a well and in a second direction in which the culture vessel moves away from the well. The perfusion culture apparatus according to this embodiment applies flow stimulation to the cells or biological tissues introduced into the culture vessels by the flow of the culture medium.

[0010] As shown in Figures 1(A) to (C), the perfusion culture apparatus 1 according to this embodiment comprises an outer container 5, a plurality of culture containers 8 (six in Figure 1(A)), a container plate 7, a base 10 on which the outer container 5 is placed, a support frame 6 positioned spaced apart from the base 10 and supporting the periphery of the container plate 7, and a drive unit 11 for moving the container plate 7 relative to the outer container 5. The outer container 5 has a flat rectangular shape in plan view and has a plurality of wells 5a (six in Figure 1(A)) opening on one side in the thickness direction. Each well 5a is, for example, circular in plan view. As shown in Figure 1(B), culture medium B1 is contained in each well 5a. The number of wells 5a is not limited to six, but may be seven or more, or five or fewer. The outer container 5 is formed from, for example, a transparent synthetic resin.

[0011] The culture vessel 8 has a cylindrical main part 801 and an outer flange part 803 that extends from the end on the +Z direction side of the main part 801 in a direction orthogonal to the axial direction of the main part 801 and away from the cylinder axis. Also, in the state where the culture vessel 8 is inserted into the well 5a of the outer container 5, the bottom side of the well, that is, the end on the -Z direction side, is closed by a porous membrane 802. As the porous membrane 802, it is preferably less than or equal to half the size of the cells or biological tissue to be cultured. For example, a membrane filter with a large number of micropores having a diameter of 0.3 μm to 10 μm can be employed. For example, when culturing liver cells, since the size of liver cells is about 20 μm, the diameter of the micropores formed in the porous membrane 802 is preferably set to 10 μm or less. Then, the cells or biological tissue to be cultured are seeded on the +Z direction side of the porous membrane 802.

[0012] As shown in FIG. 1(A), the container plate 7 has a rectangular plate-shaped main piece 701 and four extension pieces 702 that extend outward from both sides in the longitudinal direction of the main piece 701, and is disposed facing the one side where each well 5a in the outer container 5 opens, that is, the +Z direction side of the outer container 5. In the part of the main piece 701 facing each of the six wells 5a of the outer container 5, a holding hole 7a that penetrates in the thickness direction of the main piece 701 and holds the culture vessel 8 inside is provided. Here, the culture vessel 8 is inserted into the holding hole 7a from the +Z direction side of the container plate 7 to the porous membrane 802 side, and the outer flange part 803 of the culture vessel 8 is held by the container plate 7 in a state where the outer peripheral part abuts against the holding hole 7a in the container plate 7.

[0013] The base 10 is rectangular in shape, and the outer container 5 is placed on its +Z side. Ball bushings 12 are also provided at the four corners of the base 10. Furthermore, as shown in Figure 1(C), the base 10 has a recess 10a into which the tip 113a of the screw 113 of the drive unit 11, which will be described later, is fitted. The support frame 6, as shown in Figure 1(A), has an overall U-shape and consists of a long main piece 601 and two leg pieces 602 extending in the same direction, i.e., the +Y direction, from each of the ends of the main piece 601 in the longitudinal direction. The support frame 6 supports the container plate 7 with the four extending pieces 702 of the container plate 7 in contact with the +Z side of the two leg pieces 602. Furthermore, as shown in Figures 1(B) and (C), four elongated guide rods 13 are fixed to the support frame 6 at both ends in the longitudinal direction of the main piece 601 and at the ends of each of the two leg pieces 602, i.e., at the ends on the +Y direction side. Here, one end of each guide rod 13 in its longitudinal direction is welded to the support frame 6. The other end of each guide rod 13 is inserted into a ball bush 12 disposed on the base 10, so that the support frame 6 can move smoothly along the Z-axis direction. In this way, because the four guide rods 13 fixed to the support frame 6 are each inserted into ball bush 12 disposed at the four corners of the base 10, the container plate 7 supported by the support frame 6 can move vertically while maintaining a state that is substantially parallel to the direction perpendicular to the vertical direction.

[0014] The drive unit 11 includes an actuator 111, a fixing unit 112 that fixes the actuator 111 to the main piece 601 of the support frame 6, and a screw 113. The actuator 111 is a so-called linear stepping motor, which incorporates a nut (not shown) that engages with the screw 113 and a stepping motor (not shown) that rotationally drives the nut. By rotationally driving the nut, the entire actuator 111 moves along the longitudinal direction of the screw 113. The tip 113a of the screw 113 is fitted into the recess 10a of the base 10. The drive unit 11 moves the support frame 6 that supports the container plate 7 along the longitudinal direction of the screw 113, thereby alternately moving the support frame 6 in the direction approaching the base 10, i.e., the -Z direction, and the direction moving away from the base 10, i.e., the +Z direction. As a result, the drive unit 11 relatively moves the container plate 7 with respect to the outer container 5 such that the culture container 8 held by the container plate 7 alternately moves in the first direction toward the bottom of the well 5a of the outer container 5 and the second direction away from the bottom of the well 5a.

[0015] As shown in FIGS. 2(A) and (B), the drive unit 11 is capable of moving the support frame 6 that supports the container plate 7 to a first position Pos1 where a part of the culture container 8 is disposed inside the well 5a of the outer container 5 and a second position Pos2 where a part of the culture container 8 is disposed inside the well 5a and is spaced apart from the base 10 more than the first position Pos1, i.e., on the +Z direction side of the first position Pos1, in a state where the culture container 8 is held by the container plate 7.

[0016] Next, the operation of the perfusion culture apparatus 1 according to this embodiment will be described. First, the user seeds the cells or biological tissue to be cultured onto the porous membrane 802 of the culture container 8 held on the container plate 7, and then places the container plate 7 holding the culture container 8 onto the support frame 6. Here, the drive unit 11 moves the support frame 6 supporting the container plate 7 to a first position Pos1, where the lower end of the culture container 8 held on the container plate 7 is positioned inside the well 5a of the outer container 5, as shown in Figure 2(A). In this case, the porous membrane 802 on which the cells or biological tissue are seeded is immersed in the culture medium B1.

[0017] Next, as shown by arrow AR12 in Figure 2(B), the drive unit 11 moves the support frame 6 that supports the container plate 7 from the first position Pos1 to the second position Pos2, where the lower end of the culture container 8 is positioned inside the well 5a of the outer container 5 and is located on the +Z side of the first position Pos1. Even when the support frame 6 is positioned at the second position Pos2, the porous membrane 802 remains immersed in the culture medium B1. Subsequently, as shown by arrow AR11 in Figure 2(A), the drive unit 11 moves the support frame 6 again from the second position Pos2 back to the first position Pos1.

[0018] Thereafter, as shown by arrow AR12 in Figure 2(B), the drive unit 11 moves the support frame 6 from the second position Pos2 to the first position Pos1, and then moves it from the first position Pos1 to the second position Pos2, repeating this process at a predetermined cycle. When the drive unit 11 moves the support frame 6 from the second position Pos2 to the first position Pos1, as shown by arrow AR11 in Figure 2(A), a pressure difference is generated such that the pressure outside the culture vessel 8 in the culture medium B1 stored in the well 5a of the outer container 5 becomes higher than the pressure inside the culture vessel 8. As a result, as shown by arrow AR21, the culture medium B1 stored in the well 5a and outside the culture vessel 8 permeates the porous membrane 802 and flows into the inside of the culture vessel 8. Here, it is preferable for the drive unit 11 to set the movement speed of the support frame 6 to a speed close to the flow velocity of tissue fluid in a living organism.

[0019] On the other hand, when the drive unit 11 moves the support frame 6 from the first position Pos1 to the second position Pos2, as shown by arrow AR12 in Figure 2(B), a pressure difference is generated such that the pressure inside the culture vessel 8 in the culture medium B1 stored in the well 5a is higher than the pressure outside the culture vessel 8. As a result, as shown by arrow AR22, the culture medium B1 stored in the well 5a and inside the culture vessel 8 flows out to the outside of the culture vessel 8 through the porous membrane 802. In this way, the drive unit 11 can generate a reciprocating flow of culture medium B1 inside the culture vessel 8 by repeatedly moving the support frame 6 back and forth along the Z-axis. This makes it possible to uniformly provide the flow stimulus necessary for culture to the cells or biological tissues being cultured inside each of the multiple culture vessels 8. The drive unit 11 moves the support frame 6 back and forth with a period dT1, for example, as shown in Figure 3(A). Furthermore, the drive unit 11 can reciprocate the support frame 6 between a first position Pos1 and a position Pos2' located at a distance equivalent to half the distance between the first position Pos1 and the second position Pos2, as shown in Figure 3(B), thereby maintaining the flow velocity of the flow generated in the culture vessel 8 while making the flow direction switching period dT2 half of the period dT1. Moreover, if the drive unit 11 moves the support frame 6 between the first position Pos1 and the second position Pos2, as shown in Figure 3(C), for example, the flow direction switching period dT2 of the culture medium B1 flow generated in the culture vessel 8 half of the period dT1, the flow velocity of the culture medium B1 can be doubled.

[0020] As described above, according to the perfusion culture apparatus 1 of this embodiment, holding holes 7a are provided through the container plate 7 in the portions facing each of the multiple wells 5a of the outer container 5 to hold the culture containers 8 on the inside. Furthermore, the drive unit 11 moves the container plate 7 relative to the outer container 5 so that the culture containers 8 held on the container plate 7 move alternately toward the bottom of the wells 5a and toward away from the bottom of the wells. In this way, by simply moving the container plate 7 relative to the outer container 5, a pressure difference can be generated between the inside and outside of the porous membrane 802 of the multiple culture containers 8. And the culture medium can be perfused in each of the multiple culture containers 8 simultaneously. Thus, there is no need for tubes and pumps to perfuse the culture medium B1 in the wells 5a of the outer container 5 and in the culture containers 8, and there is an advantage that cell or biological tissue perfusion culture can be performed with a simple configuration consisting of a container plate 7 and a drive unit 11 that drives the container plate 7. Furthermore, since there is no need to continuously supply medium B1 from an external source during perfusion culture of cells or biological tissues, contamination of medium B1 with microorganisms and other foreign substances during external supply can be suppressed.

[0021] Incidentally, a perfusion culture apparatus has been proposed that comprises an outer container which is flat and box-shaped with one side open in the thickness direction and stores culture medium B1 inside; a container plate supported by the side wall of the outer container which holds multiple culture containers 8 with the porous membrane 802 side of each container immersed in the culture medium B1 stored in the outer container; and a movable plate driving mechanism which swings a movable plate to which the outer container is fixed, with its center as the pivot point. This perfusion culture apparatus generates flow in the culture medium B1 stored in the outer container 5 by swinging the movable plate. However, in the case of this perfusion culture apparatus, the flow velocity of the culture medium B1 becomes uneven in the outer container, making it difficult to provide a uniform flow stimulus to the cells or biological tissues placed in the multiple culture containers 8. Furthermore, in the case of this perfusion culture apparatus, the flow of culture medium B1 mainly occurs in a direction perpendicular to the thickness direction of the porous membrane 802 of the culture container 8, making it difficult to generate the flow of culture medium B1 in the thickness direction of the porous membrane 802 that is required, for example, in the culture of liver cells or liver tissue. In contrast, the perfusion culture apparatus 1 according to this embodiment can move multiple culture vessels 8 at the same speed in the Z-axis direction, thereby making the flow rate of the culture medium B1 uniform among the multiple culture vessels 8. Therefore, it is possible to suppress the occurrence of unevenness in the culture conditions among the cells or biological tissues introduced into each of the multiple culture vessels 8. Furthermore, by moving the culture vessels 8 and the vessel plate 7 in the Z-axis direction, the perfusion culture apparatus 1 can generate a flow of culture medium B1 mainly in the thickness direction of the porous membrane 802, thereby realizing a culture environment suitable for culturing liver cells or liver tissue, for example.

[0022] (Embodiment 2) The perfusion culture system according to this embodiment comprises a perfusion culture apparatus described in Embodiment 1, a housing that encloses the perfusion culture apparatus, a temperature measuring unit for measuring the temperature inside the housing, a heater for heating the inside of the housing, and a control device that controls the heater to maintain the temperature inside the housing at a preset temperature based on the temperature measured by the temperature measuring unit.

[0023] The perfusion culture system according to this embodiment, as shown in Figure 4, for example, comprises a perfusion culture apparatus 1 described in Embodiment 1, a housing 21 that houses the perfusion culture apparatus 1 inside, a temperature measuring unit 23 for measuring the temperature inside the housing 21, a heater 22, and a control device 29. In Figure 4, components similar to those in Embodiment 1 are denoted by the same reference numerals as in Figures 1(A) to (C). The housing 21 is formed in the shape of a rectangular box from a material with relatively low thermal conductivity. Here, a transparent polycarbonate resin can be used as the material with relatively low thermal conductivity. The heater 22 is, for example, a resistance heater and is arranged at multiple locations (four locations in Figure 4) on the inner wall of the housing 21. The temperature measuring unit 23 is, for example, a temperature sensor using a thermistor and is arranged one at a time near each of the locations on the inner wall of the housing 21 where the multiple heaters 22 are arranged.

[0024] The perfusion culture system further includes a humidity measuring unit 281 provided inside the housing 21 to measure the humidity inside the housing 21, and a steam supply source 31 to supply steam into the housing 21. The steam supply source 31 has a storage unit 311 provided on the inner wall of the housing 21 to store liquid, and a heating unit 312 that supplies steam into the housing 21 by heating the liquid stored in the storage unit 311.

[0025] Furthermore, the perfusion culture system includes a tube PI1 communicating with the housing 21, a first tank 241 communicating with tube PI1 via tube PI2 for storing carbon dioxide gas, and a second tank 251 communicating with tube PI1 via tube PI3 for storing nitrogen gas. The perfusion culture system also includes a valve 26 provided at the outside air inlet of tube PI1, a filter 27 inserted at the inlet of tube PI1 to the housing 21, and a gas concentration measuring unit 282 provided inside the housing 21 for measuring the concentrations of carbon dioxide and nitrogen. Control valves 242 and 252 are interposed in tubes PI2 and PI3, respectively, for adjusting the amount of carbon dioxide gas or nitrogen gas supplied from the first tank 241 and the second tank 251 to tube PI1. Here, a gas supply unit is configured to supply carbon dioxide gas into the housing 21 from the first tank 241 and control valve 242, and a gas supply unit is configured to supply nitrogen gas into the housing 21 from the second tank 251 and control valve 252. In addition, a pipe PI4 that communicates with the housing of another perfusion culture apparatus (not shown) is connected to pipe PI1, so that cells or biological tissues can be cultured simultaneously in multiple perfusion culture systems.

[0026] The control device 29 controls the heaters 22 to maintain the temperature inside the housing 21 at a preset temperature, based on the temperature measured by the temperature measuring unit 23. Here, the control device 29 controls each of the multiple heaters 22 individually based on the temperature measured by the corresponding temperature measuring unit 23. The control device 29 also controls the heating unit 312 of the steam supply source 31 to maintain the humidity measured by the humidity measuring unit 281 at a preset humidity. Furthermore, the control device 29 controls the amount of gas supplied from the first tank 241 and the second tank 251 into the housing 21 to maintain the concentrations of carbon dioxide gas and nitrogen gas inside the housing 21 at a preset concentration, based on the concentrations of carbon dioxide gas and nitrogen gas measured by the gas concentration measuring unit 282. Specifically, the control device 29 adjusts the flow rates of carbon dioxide gas and nitrogen gas flowing from the first tank 241 and the second tank 251 to the pipe PI1 by controlling the opening degrees of the control valves 242 and 252, respectively, which correspond to the first tank 241 and the second tank 251. The control device 29 also controls the open and closed state of the valve 26. When opening the housing 21 to the atmosphere, the control device 29 introduces outside air into the housing 21 by changing the valve 26 from the closed state to the open state.

[0027] As described above, according to the perfusion culture system of this embodiment, the temperature inside the housing 21 is maintained at a preset temperature based on the temperature measured by the temperature measurement unit 23. This suppresses the occurrence of convection caused by temperature fluctuations inside the culture vessel 8 of the perfusion culture device 1, thereby enabling stable perfusion culture of cells or biological tissues inside the culture vessel 8.

[0028] Furthermore, according to the perfusion culture system of this embodiment, the humidity measured by the humidity measurement unit 281 is maintained at a preset humidity, and the concentrations of carbon dioxide gas and nitrogen gas measured by the gas concentration measurement unit 282 are maintained at preset concentrations. This has the advantage of creating a culture environment equivalent to that of an incubator inside the housing 21. Moreover, because the housing 21 is transparent, it is possible to culture cells or biological tissues while continuously monitoring the inside of the culture container 8 of the perfusion culture device 1 with an optical microscope from, for example, the +Z direction side of the housing 21.

[0029] (Embodiment 3) The perfusion culture system according to this embodiment comprises a perfusion culture apparatus described in Embodiment 1, a cassette having a plurality of slots arranged vertically, in which an outer container used in the perfusion culture apparatus is placed in at least one of the plurality of slots, and a transport device for transporting the outer container from either the perfusion culture apparatus or the cassette to the other. This perfusion culture system automates the setting of the outer container into the perfusion culture apparatus and is useful for continuously performing culture processing on a large number of samples, such as in drug discovery screening.

[0030] As shown in Figure 5, the perfusion culture system according to this embodiment comprises a perfusion culture device 1, a cassette 3030 for storing the outer container 5, and a transport device 3040 for transporting the outer container 5 from either the perfusion culture device 1 or the cassette 3030 to the other. In Figure 5, components similar to those in Embodiment 1 are denoted by the same reference numerals as in Figures 1(A) to (C). The perfusion culture system also comprises a housing 3021 that houses the perfusion culture device 1, the cassette 3030, and the transport device 3040, an analyzer 3075 for analyzing the culture medium stored in the wells 5a of the outer container 5, and a gate 3050 interposed between the housing 3021 and the analyzer 3075. Furthermore, as shown in Figure 6, the perfusion culture system includes a reagent injection unit 3060 equipped with a nozzle 3061 for injecting reagents into the culture vessel 8, a microscope unit 3070 for observing the culture medium stored in the wells 5a of the outer container 5, a plate moving unit 3080 for moving the container plate 7 of the perfusion culture apparatus 1 in the Z-axis direction, and a support stand 3090 for supporting the perfusion culture apparatus 1. Also, as shown in Figure 5, the perfusion culture system includes a control device 3029 for controlling the perfusion culture apparatus 1, a transport device 3040, an analyzer 3075, a gate 3050, a reagent injection unit 3060, a microscope unit 3070, and a plate moving unit 3080.

[0031] The housing 3021 is box-shaped, and a gate 3050 for inserting the outer container 5 into the analyzer 3075 is provided on a part of its peripheral wall. The housing 3021 has, for example, a gas introduction section (not shown) for introducing gas into the housing 3021, and a purification filter (not shown) for purifying the introduced gas, and prevents outside air from flowing into the housing 3021 by maintaining positive pressure inside the housing 3021. The analyzer 3075 is an instrument for analyzing culture medium B1. The analyzer 3075 transmits analysis result information indicating the analysis results to the control device 3029.

[0032] The cassette 3030 has multiple (four in Figure 7) slots SL arranged vertically, i.e., in the Z-axis direction, as shown in Figure 6, for example, and the outer container 5 is placed in the slots SL. Here, the cassette 3030 has two support parts 3031 arranged side by side in the X-axis direction, and multiple ribs 3032 protruding from opposite sides of each of the two support parts 3031. The two support parts 3031 and the pair of opposing ribs 3032 provided on each of the two support parts 3031 form a slot SL on the +Z direction side of the pair of ribs 3032 on which the lower end of the periphery of the outer container 5 is placed.

[0033] As shown in Figure 5, the transport device 3040 includes a transport robot 3041 and a rail 3042 laid along the X-axis direction on which the transport robot 3041 moves. When the transport robot 3041 transports the outer container 5 used for culture in the perfusion culture device 1 to the cassette 3030, it first moves to a position on the rail 3042 facing the perfusion culture device 1. Next, the transport robot 3041 extends its arm 3041a to receive the outer container 5 from the perfusion culture device 1, and then retracts the arm 3041a. After that, with the outer container 5 held in the arm 3041a, the transport robot 3041 moves to a position on the rail 3042 facing the cassette 3030, and then extends the arm 3041a to place the outer container 5 in the slot SL of the cassette 3030. Furthermore, when the transport robot 3041 transports the outer container 5 stored in the cassette 3030 to the perfusion culture apparatus 1, it first moves to a position on the rail 3042 facing the cassette 3030. Next, the transport robot 3041 extends its arm 3041a to receive the outer container 5 from the cassette 3030, and then retracts the arm 3041a. After that, the transport robot 3041 moves to a position on the rail 3042 facing the perfusion culture apparatus 1 while holding the outer container 5 in its arm 3041a, and then extends its arm 3041a to place the outer container 5 on the base 10 of the perfusion culture apparatus 1. In addition, if the gate 3050 is open, the transport robot 3041 extends the arm 3041a holding the outer container 5 through the gate 3050 toward the inside of the analyzer 3075, thereby passing the outer container 5 to the analyzer 3075.

[0034] As shown in Figure 6, the reagent injection unit 3060 is positioned opposite the perfusion culture apparatus 1 on the +Z side of the perfusion culture apparatus 1. The reagent injection unit 3060 has the aforementioned plurality of nozzles 3061 and a head 3062 that supplies reagents to the nozzles 3061. The microscope unit 3070 has a microscope 3071 having a camera and positioned opposite the perfusion culture apparatus 1 or cassette 3030 on the +Z side of the perfusion culture apparatus 1 and cassette 3030, a microscope drive unit 3072 that moves the microscope 3071 in the X-axis or Y-axis direction, and a microscope unit control unit (not shown) that transmits image information showing the image captured by the microscope 3071 to the control device 3029. Here, the microscope drive unit 3072 is capable of moving the microscope 3071 in the XY direction on the +Z side of the perfusion culture apparatus 1 or cassette 3030. As a result, the microscope 3071 can use its camera to image the culture medium B1 stored inside each of the multiple culture vessels 8 of the perfusion culture apparatus 1, or inside each of the multiple wells 5a of the outer container 5 located on the top row of the cassette 3030. By equipping the perfusion culture system with such a microscope unit 3070, the user can observe the morphology or density of cells present in the culture vessels 8 or the wells 5a of the outer container 5. The plate moving unit 3080 has a chuck 3081 that holds the container plate 7 of the perfusion culture apparatus 1, and a chuck drive unit 3082 that moves the chuck 3081 relative to the support frame 6 of the perfusion culture apparatus 1. The chuck 3081 is, for example, a vacuum chuck. The chuck drive unit 3082 moves the chuck 3081 holding the container plate 7 toward the +Z direction, thereby detaching the container plate 7 from the support frame 6.

[0035] The control device 3029 is, for example, a programmable logic controller and includes a memory (not shown) that stores a program for performing a culture process to continuously culture cells or biological tissues, and a CPU unit (not shown) that reads and executes the program stored in the memory. The control device 3029 also includes storage (not shown) that stores image information transmitted from the microscope unit 3070 and analysis result information transmitted from the analyzer 3075. Furthermore, the control device 3029 controls the operation of the perfusion culture apparatus 1, the transport apparatus 3040, the analyzer 3075, the gate 3050, the reagent injection unit 3060, the microscope unit 3070, and the plate moving unit 3080 individually by outputting control signals to each of them. In addition, the control device 3029 has a user interface (not shown) with a display device and an input device. Here, the display device is, for example, a liquid crystal display, and the input device is a keyboard, touchpad, etc.

[0036] Next, the culture process performed by the perfusion culture system according to this embodiment will be described with reference to Figures 7 to 12. This culture process is initiated when power is supplied to the perfusion culture system. The plate moving unit 3080 is assumed to have moved the chuck 3081 that holds the container plate 7 to a position where the container plate 7 is detached from the support frame 6 when the power is supplied. First, the control device 3029 determines whether a processing start command has been issued by the user performing an operation to start the culture process via the user interface (step S101). The control device 3029 repeatedly executes the process in step S101 unless a processing start command has been issued (step S101: No).

[0037] Meanwhile, when the control device 3029 determines that a processing start command has been issued (step S101: Yes), it outputs a control signal to the transport device 3040 to transport the outer container 5 from the cassette 3030 to the perfusion culture device 1. When the control signal is input to the transport device 3040, the transport robot 3041 transports one of the outer containers 5 stored in the cassette 3030 to the perfusion culture device 1 (step S102). Here, the transport robot 3041 receives the outer container 5 placed in the slot SL located at the bottom of the cassette 3030 and then moves to a position on the rail 3042 facing the perfusion culture device 1. Then, the transport robot 3041 extends its arm 3041a to place the outer container 5 on the base 10 of the perfusion culture device 1.

[0038] Next, the plate moving unit 3080 places the container plate 7 on the support frame 6 by moving the chuck 3081 that holds the container plate 7 in the -Z direction based on a control signal input from the control device 3029 (step S103). At this time, the lower end of the culture container 8 holding the container plate 7 is immersed in the culture medium B1 stored in the well 5a of the outer container 5. Subsequently, the drive unit 11 of the perfusion culture apparatus 1 starts a repeated lifting and lowering operation based on a control signal input from the control device 3029 (step S104). Here, "repeated lifting and lowering operation" means that the drive unit 11 moves the support frame 6 from the second position Pos2 described in Embodiment 1 to the first position Pos1, and then moves it from the first position Pos1 to the second position Pos2, repeating this operation at a preset period. The control device 3029 also appropriately changes the repetition period and the movement width of the support frame 6 in the repeated lifting and lowering operation based on the analysis result information transmitted from the analyzer 3075, which will be described later.

[0039] Subsequently, the control device 3029 determines whether or not a preset observation period has arrived (step S105). This observation period is set, for example, based on the time from when the drive unit 11 starts its repeated up-and-down movement until the surface of the cells or biological tissue in the culture vessel 8 rises to near the top surface of the culture medium B1. If the control device 3029 determines that the observation period has not yet arrived (step S105: No), the process described in step S113 below is executed. On the other hand, if the control device 3029 determines that the observation period has arrived (step S105: Yes), it outputs a control signal to the drive unit 11 to stop the repeated up-and-down movement of the support frame 6 by the drive unit 11 (step S106). Next, the plate moving unit 3080 detaches the container plate 7 from the support frame 6 based on the control signal input from the control device 3029 (step S107). At this time, the plate moving unit 3080 moves the chuck 3081 that holds the container plate 7 in the +Z direction, for example as shown by arrow AR31 in Figure 8(A), thereby detaching the container plate 7 from the support frame 6.

[0040] Returning to Figure 7, the transport robot 3041 of the transport device 3040 then transports the outer container 5, which is placed on the base 10 of the perfusion culture device 1, to the uppermost slot SL of the cassette 3030 based on the control signal input from the control device 3029 (step S108). At this time, the transport robot 3041 moves to a position on the rail 3042 facing the perfusion culture device 1, and then receives the outer container 5 placed on the base 10 of the perfusion culture device 1, as shown by arrow AR32 in Figure 8(B). Then, the transport robot 3041 moves to a position on the rail 3042 facing the cassette 3030, as shown by arrow AR33 in Figure 8(B). After that, the transport robot 3041 places the outer container 5 in the uppermost slot SL of the cassette 3030 by extending its arm 3041a, as shown by arrow AR34 in Figure 9(A).

[0041] Returning to Figure 7, the microscope unit 3070 then performs an observation process based on a control signal input from the control device 3029, which involves imaging the culture medium B1 stored inside each of the multiple culture vessels 8 of the perfusion culture apparatus 1, or in the wells 5a of the outer container 5, generating image information showing the captured image, and transmitting it to the control device 3029 (step S109). Here, the microscope unit 3070, for example as shown by arrow AR35 in Figure 9(B), uses the microscope drive unit 3072 to move the microscope 3071 while imaging the culture medium B1 stored inside each of the multiple culture vessels 8 of the perfusion culture apparatus 1, or in each of the multiple wells 5a of the outer container 5. Returning to Figure 7, the transport robot 3041 of the transport device 3040 then transports the outer container 5, which is placed in the uppermost slot SL of the cassette 3030, to the perfusion culture apparatus 1 based on a control signal input from the control device 3029 (step S110). At this point, the transport robot 3041 moves to a position on the rail 3042 facing the cassette 3030, and then receives the outer container 5 placed in the uppermost slot SL of the cassette 3030, as shown by arrow AR36 in Figure 10(A). Then, the transport robot 3041 moves to a position on the rail 3042 facing the perfusion culture device 1, as shown by arrow AR37 in Figure 10(A). After that, the transport robot 3041 extends its arm 3041a, as shown by arrow AR38 in Figure 10(A), to place the outer container 5 on the base 10 of the perfusion culture device 1.

[0042] Returning to Figure 7, the plate moving unit 3080 then places the container plate 7 onto the support frame 6 based on a control signal input from the control device 3029 (step S111). At this time, the plate moving unit 3080 places the container plate 7 onto the support frame 6 by moving the chuck 3081 that holds the container plate 7 in the -Z direction, for example as shown by arrow AR39 in Figure 10(B). Returning to Figure 7, the drive unit 11 of the perfusion culture apparatus 1 then starts the repeated lifting and lowering operation again based on a control signal input from the control device 3029 (step S112).

[0043] Subsequently, the control device 3029 determines whether or not the preset medium exchange time has arrived (step S113). This medium exchange time is set to be an integer multiple of the time from when the drive unit 11 starts its repeated up-and-down movement until the surface of the cells or biological tissue in the culture vessel 8 rises to near the top surface of the medium B1. If the control device 3029 determines that the medium exchange time has not yet arrived (step S113: No), the process in step S128, described later, is executed. On the other hand, if the control device 3029 determines that the medium exchange time has arrived (step S113: Yes), it outputs a control signal to the drive unit 11 to stop the repeated up-and-down movement of the support frame 6 by the drive unit 11 (step S114). Next, the plate moving unit 3080 detaches the container plate 7 from the support frame 6 based on the control signal input from the control device 3029 (step S115). Next, the transport robot 3041 of the transport device 3040 transports the outer container 5, which is placed on the base 10 of the perfusion culture device 1, to the uppermost slot SL of the cassette 3030 based on the control signal input from the control device 3029 (step S116). After that, the transport robot 3041 of the transport device 3040 transports the outer container 5, which is placed in the lowermost slot SL of the cassette 3030, to the perfusion culture device 1 based on the control signal input from the control device 3029 (step S117).

[0044] Next, as shown in Figure 11, the plate moving unit 3080 places the container plate 7 onto the support frame 6 based on a control signal input from the control device 3029 (step S118). Subsequently, the drive unit 11 of the perfusion culture apparatus 1 starts the repeated lifting and lowering operation again based on a control signal input from the control device 3029 (step S119).

[0045] Subsequently, the control device 3029 determines whether or not the device is set to perform analysis of the culture medium B1 stored in well 5a of the outer container 5 using the analyzer 3075 (step S120). The control device 3029 determines whether or not the device is set to perform analysis of the culture medium B1 stored in well 5a of the outer container 5 using the analyzer 3075 (step S120). If the control device 3029 determines that the device is not set to perform analysis using the analyzer 3075 (step S120: No), the process in step S128 described below is executed. On the other hand, if the control device 3029 determines that the device is set to perform analysis using the analyzer 3075 (step S120: Yes), it outputs a control signal to the gate 3050 to open the gate 3050. As a result, the gate 3050 opens (step S121).

[0046] Next, the transport robot 3041 of the transport device 3040 transports the outer container 5, which is located in the uppermost slot SL of the cassette 3030, to the analyzer 3075 based on the control signal input from the control device 3029 (step S122). Here, when the transport robot 3041 receives the outer container 5 from the cassette 3030, it rotates its arm 3041a as shown by arrow AR40 in Figure 12(A). Then, the transport robot 3041 extends its arm 3041a as shown by arrow AR41 in Figure 12(B), thereby passing the outer container 5 to the analyzer 3075. Returning to Figure 11, the control device 3029 then outputs a control signal to the gate 3050, thereby closing the gate 3050 (step S123). Subsequently, the analyzer 3075 performs the analysis of the culture medium B1 stored in the well 5a of the outer container 5 based on the control signal input from the control device 3029 (step S124). Then, when the analysis process of the analysis device 3075 is completed, the control device 3029 outputs a control signal to the gate 3050, thereby opening the gate 3050 (step S125). Next, the transport robot 3041 of the transport device 3040 transports the outer container 5 placed inside the analysis device 3075 to the uppermost slot SL of the cassette 3030 based on the control signal input from the control device 3029 (step S126). Subsequently, the control device 3029 outputs a control signal to the gate 3050, thereby closing the gate 3050 again (step S127). After that, the transport robot 3041 of the transport device 3040 rearranges the outer containers 5 stored in the cassette 3030 by moving the outer containers 5 placed in each slot SL of the cassette 3030 to the next lower level based on the control signal input from the control device 3029 (step S128).

[0047] Subsequently, the control device 3029 determines whether or not the preset reagent injection time has arrived (step S129). If the control device 3029 determines that the reagent injection time has not yet arrived (step S129: No), the process in step S135 described later is executed. On the other hand, if the control device 3029 determines that the reagent injection time has arrived (step S129: Yes), it outputs a control signal to the drive unit 11, thereby stopping the repeated raising and lowering operation of the support frame 6 by the drive unit 11 (step S130). Next, the reagent injection unit 3060 lowers the head 3062 from the standby position based on the control signal input from the control device 3029 (step S131). Subsequently, the reagent injection unit 3060 injects the reagent into the culture vessel 8 through the nozzle 3061 based on the control signal input from the control device 3029 (step S132). Subsequently, the reagent injection unit 3060 raises the head 3062 to the standby position based on the control signal input from the control device 3029 (step S133). Next, the drive unit 11 of the perfusion culture apparatus 1 starts the repeated raising and lowering operation again based on the control signal input from the control device 3029 (step S134).

[0048] Next, the control device 3029 determines whether or not a processing termination command has been issued to end the culture process (step S135). This processing termination command is issued, for example, when the culture process is completed for all outer containers 5 placed in the cassette 3030. If the control device 3029 determines that a processing termination command has not yet been issued (step S135: No), the process in step S102 is executed again. On the other hand, if the control device 3029 determines that a processing termination command has been issued (step S135: Yes), the culture process is terminated.

[0049] As described above, the perfusion culture system according to this embodiment allows for the automatic replacement of culture medium B1, making it possible to efficiently culture cells or biological tissues.

[0050] Although various embodiments of the present invention have been described above, the present invention is not limited to the configurations of the embodiments described above. For example, as shown in Figure 13, the perfusion culture apparatus 4001 may have a vibration-damping member 4019 interposed between the fixed part 112 of the drive unit 11 and the support frame 6. As the vibration-damping member 4019, for example, a gel-like substance mainly composed of silicone resin can be used.

[0051] This configuration has the advantage of suppressing the generation of convection in the culture medium B1 inside the culture vessel 8 caused by vibrations emitted from the actuator 111, thereby enabling proper culture of cells or biological tissues.

[0052] In Embodiment 1, for example, as shown in Figure 14, a fixation material 5804 for fixing cells or biological tissue onto the porous membrane 802 may be placed on the porous membrane 802 side of the culture vessel 8. Preferably, the fixation material 5804 is a substance that does not have a chemical or physical effect on the cells or biological tissue and undergoes a phase change within a processing temperature range. Preferably, the fixation material 5804 is agarose, for example, which undergoes a phase change from liquid to solid at 37°C and allows the culture medium B1 to permeate in the solid phase. First, cells or biological tissue are suspended in the liquid phase of the fixation material 5804 and seeded onto the porous membrane 5802 with the cells or biological tissue uniformly diffused within the fixation material 5804. Then, the temperature of the culture medium B1 is lowered to 37°C to cause the fixation material 5804 to undergo a phase transition to the solid phase. At this time, the fixation material 5804 is adhered to the inner wall of the culture vessel 8 and the porous membrane 802.

[0053] However, if the fixation material 5804 is not used, cells or biological tissues may adhere to the porous membrane 802, potentially becoming an obstacle to the flow of culture medium B1 through the porous membrane 802. Alternatively, cells or biological tissues attached to the porous membrane 802 may detach from the porous membrane 802 due to the flow of culture medium B1 within the culture vessel 8, and end up floating on the uppermost surface of the culture medium B1. In contrast, with this configuration, the fixation material 5804 maintains the cells or biological tissues immersed in the culture medium B1 within the culture vessel 8, thus allowing the cells or biological tissues to be appropriately stimulated by the flow of culture medium B1.

[0054] Embodiment 1 describes an example of a perfusion culture apparatus 1 equipped with a ball bush 12. However, the perfusion culture apparatus is not limited to this, and may, for example, be equipped with a cylindrical member (not shown) in which the gap between the side wall and the inner wall of the guide rod 13 is extremely small instead of the ball bush 12. Furthermore, the drive unit may have an actuator having a piston fixed to the support frame 6, a cylinder into which the piston is inserted, and a fluid supply unit that supplies fluid into the cylinder. Moreover, the drive unit may have a plurality of actuators that drive the support frame 6. In this case, the plurality of actuators operate synchronously with respect to each other, thereby maintaining a state that is substantially parallel to a plane perpendicular to the vertical direction of the support frame 6.

[0055] In Embodiment 2, an example was described in which the housing 21 is formed in the shape of a rectangular box from, for example, a transparent polycarbonate resin. However, the invention is not limited to this, and for example, the housing 21 may have a structure in which a vacuum insulation layer is formed over substantially the entire surface of its peripheral wall. Alternatively, the housing may be box-shaped, with only a portion of the peripheral wall being made of a transparent material.

[0056] Although embodiments and variations of the present invention have been described above, the present invention is not limited thereto. The present invention includes embodiments and variations that are appropriately combined, and those that are appropriately modified thereto. [Industrial applicability]

[0057] This invention is suitable for techniques that involve simultaneously culturing cells or biological tissues in multiple culture vessels. [Explanation of Symbols]

[0058] 1,4001: Perfusion culture apparatus, 5: Outer container, 5a: Well, 6: Support frame, 7: Container plate, 7a: Holding hole, 8: Culture vessel, 10: Base, 10a: Recess, 11: Drive unit, 12: Ball bush, 13: Guide rod, 21,3021: Housing, 22: Heater, 23: Temperature measuring unit, 26: Valve, 27: Filter, 29,3029: Control device, 31: Steam supply source, 111: Actuator, 112: Fixing unit, 113: Screw, 113a: Tip, 241: First tank, 242,252: Control valve, 251: Second tank, 281: Humidity measuring unit, 282: Gas concentration measuring unit, 311: Storage unit, 312: Heating unit, 601,701: Main piece, 602: Leg 702: Extension piece, 801: Main part, 802: Porous membrane, 803: Outer flange, 3030: Cassette, 3031: Support part, 3032: Rib, 3040: Conveying device, 3041: Conveying robot, 3041a: Arm, 3042: Rail, 3050: Gate, 3060: Reagent injection unit, 3061: Nozzle, 3062: Head, 3070: Microscope unit, 3071: Microscope, 3072: Microscope drive unit, 3075: Analytical device, 3080: Plate moving part, 3081: Chuck, 3090: Support base, 4019: Vibration damping member, 5804: Fixing material, B1: Culture medium, PI1, PI2, PI3, PI4: Tube, Pos1: First position, Pos2: Second position, SL: Slot

Claims

1. an outer container having a plurality of wells for containing a culture medium; a plurality of cylindrical culture vessels, the bottom ends of which are closed with porous membranes onto which cells or biological tissues are seeded when the vessels are inserted into the wells; a plate-shaped container plate that is disposed opposite the opening of each of the plurality of wells in the outer container, and has holding holes formed in portions facing each of the plurality of wells to hold the culture containers on the inside; a drive unit that moves at least one of the container plate and the outer container relative to the other so that at least one of the culture containers held by the container plate alternates between a first direction toward the bottom of the well and a second direction away from the bottom of the well; Perfusion culture device.

2. a base on which the outer container rests; a support frame spaced apart from the base and supporting a periphery of the container plate; the driving unit alternately moves the support frame supporting the container plate in a direction toward the base and a direction away from the base, thereby alternately moving the container plate in the first direction and the second direction. The perfusion culture apparatus according to claim 1.

3. a plurality of cylindrical culture vessels, the plurality of wells of which, when inserted into the wells, have bottom ends closed with porous membranes onto which cells or biological tissues are seeded; a plate-like vessel plate, the plurality of wells of which are disposed opposite the openings of the outer vessel, and the vessel plate having holding holes formed therethrough in portions facing the wells for holding the culture vessels; and a drive unit that moves at least one of the vessel plate and the outer vessel relative to the other so that at least one culture vessel held in the vessel plate alternates between a first direction toward the bottom of the well and a second direction away from the bottom of the well; A housing that houses the perfusion culture device inside; a temperature measuring unit that measures the temperature inside the housing; a heater that heats the inside of the housing; a control device that controls the heater based on the temperature measured by the temperature measuring unit so that the temperature inside the housing is maintained at a preset temperature, Perfusion culture system.

4. a humidity measuring unit provided in the housing and configured to measure the humidity inside the housing; a steam supply source that supplies steam into the housing, the control device controls the steam supply source so that the humidity measured by the humidity measuring unit is maintained at a preset humidity. The perfusion culture system according to claim 3 .

5. a gas supply unit that supplies a preselected type of gas into the enclosure; a gas concentration measuring unit provided in the housing and configured to measure the concentration of the gas; the control device controls the amount of gas supplied from the gas supply unit into the housing based on the concentration of the gas measured by the gas concentration measurement unit so that the concentration of the gas in the housing is maintained at a preset concentration. The perfusion culture system according to claim 3 or 4.

6. The housing is box-shaped, and at least a part of the peripheral wall is formed from a transparent material. The perfusion culture system according to any one of claims 3 to 5.

7. a plurality of cylindrical culture vessels, the bottom ends of which are closed with porous membranes onto which cells or biological tissues are seeded when the vessels are inserted into the wells; a plate-like vessel plate, the vessel plate being disposed opposite the opening of each of the plurality of wells in the outer vessel, and having holding holes formed in portions facing each of the plurality of wells to hold the culture vessels; and a drive unit that moves at least one of the vessel plate and the outer vessel relative to the other so that at least one of the culture vessels held in the vessel plate moves alternately in a first direction toward the bottom of the well and a second direction away from the bottom of the well; a cassette having a plurality of vertically aligned slots, the outer container being disposed in at least one of the plurality of slots; a transport device that transports the outer container from one of the perfusion culture device and the cassette to the other; Perfusion culture system.

8. The perfusion culture apparatus includes: a base on which the outer container is placed; a support frame spaced from the base and supporting a periphery of the container plate; the driving unit alternately moves the support frame supporting the container plate in a direction toward the base and a direction away from the base, thereby alternately moving the container plate in the first direction and the second direction. The perfusion culture system according to claim 7.

9. The driving unit can position a support frame supporting the vessel plate, with the culture vessel held on the vessel plate, at either a first position where a portion of the culture vessel is disposed inside the well, or a second position where a portion of the culture vessel is disposed inside the well and is farther from the base than the first position, and moves the support frame from the second position to the first position, and then from the first position to the second position, repeatedly, thereby moving the vessel plate alternately in the first direction and the second direction. The perfusion culture system according to claim 8.

10. a plate moving unit including a chuck for holding the container plate and a chuck driving unit for moving the chuck relative to the support frame; the conveying device causes the plate moving unit to move the chuck holding the container plate in a direction away from the support frame, thereby removing the outer container placed on the base with the culture container positioned outside the well. The perfusion culture system according to claim 8 or 9.

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