JP6535181B2 - Cell administration device - Google Patents

Description

  The present invention relates to a cell administration device for seeding cells on a porous cell support.

A small amount of cartilage is collected from the patient's ear, grown and cultured, and the cultured chondrocytes are seeded on a porous structure support to obtain an implant-type regenerated cartilage, which is used as the implant-type regenerated cartilage in the patient's body. Methods for treating, for example, cartilage deformities and defects in the face by implanting in the lung are being studied.
In order to promote tissue formation while controlling cell growth and differentiation, the cells are seeded on a porous structure support, but filling the medium with the suspended cell up to the central part of the porous three-dimensional structure It was difficult.
Therefore, Patent Document 1 has the communication small hole structure having a hole diameter of 5 to 3200 μm and an average hole diameter of 50 to 1500 μm, and is guided in the entire communication small hole structure of a sheet or block support having a thickness of 2 mm or more. After filling the aqueous solution, the culture solution in which the cells are suspended is supplied to the upper surface of the support, and the induced aqueous solution is aspirated from the lower surface of the support, and the induced aqueous solution sucked from the lower surface of the support and the upper surface of the support A method is disclosed in which a negative pressure is generated between the supplied cell suspension and the supplied cell suspension is supplied into the support.

JP, 2009-195682, A

In this prior art, the support is immersed in a DMEM culture solution, which is an induction aqueous solution, placed in a sterilized desiccator, and then the pressure in the desiccator is reduced by a vacuum pump to infiltrate the induction aqueous solution into the inside of the support. There is disclosed a method of transferring the prepared transplanted cells in the culture medium by transferring them onto a 7 mm paper-based sterile filter paper moisture absorber.
However, in the prior art, there is no disclosure as to what means to carry out for the supply of the induction aqueous solution and the suspension to the support and the suction of the induction aqueous solution. In addition, although it is necessary to simultaneously prepare a plurality of supports supplied with the cell suspension in consideration of aseptic tests and tests based on the pharmacopoeia, the suspension is supplied uniformly in manual production. Although it is difficult to obtain a support, the prior art only discloses a method for seeding cells on a single support, and the method for seeding regenerated chondrocytes on a plurality of supports is disclosed. It has not been.
Therefore, the present invention aims to provide a cell administration device suitable for providing a large amount of support in the future, because the use thereof is expected to be expanded.

In order to solve the above problems, the invention according to claim 1 has a support housing recess formed on the upper surface, and a slit passing through the bottom wall of the support housing recess. A cell administration container for containing a porous cell support in a support containing recess, and a cell suspension feeder for supplying a cell suspension to the porous cell support contained in the support containing recess; And a water absorbing material contacting means for bringing a water absorbing material for absorbing water through the slit into and out of the slit from the lower side of the induction aqueous solution impregnated in the porous cell support.
The invention according to claim 2, the volume of the support housing recess, at least the porous cell support volume and the porous cell support sum and whether such a volume of liquid which can be impregnated in, It is characterized by being larger than this.

In the invention according to claim 3, the porous cell support has a communicating small pore structure having an average pore diameter of 200 to 300 μm, and is provided with a curved surface which is partially bulged outward in a dome shape. The inner surface of the support housing recess has a curved shape that is in surface contact with at least a part of the curved surface when the porous support is housed with the curved surface as the lower surface .
The invention according to claim 4 is characterized in that the support housing recess has a curved inner surface which is convex downward, and the slit is formed at a position corresponding to the top of the inner surface.
The invention according to claim 5 is characterized in that a plurality of the cell administration containers, a plurality of the water absorbing material contacting means provided corresponding to each of the cell administration containers, and a plurality of the cell suspension feeders are provided. And a cell suspension feeder driving means driven across the cell administration container.

  According to the present invention, it is possible to mechanize the production of a cell-seeded support, so that it is possible to produce a homogeneous quality support in a large amount in a short time.

It is a three-sided figure which shows the whole cell administration apparatus. It is a figure which shows an example of a porous cell support body. It is a figure which shows an example of a cell administration container. It is a figure which shows an example of a container cradle. It is a figure which shows an example of a stabilizer. It is a front view of a syringe. It is a perspective view which shows an example of a cell mixer. It is a three-sided figure which shows a cell administration container installation part and a water absorbing material contact / separation means. It is a three-sided view showing a cell suspension feeder. It is a figure which shows a water absorbing material holding member, (a) is a three-view figure which shows a water absorbing material holding base, (b) is a top view which shows a pressing frame, (c) is a water absorbing material holding holding water absorbing material. It is a sectional view of a member. It is a figure showing an example of a cushion material. It is a perspective view of a cartridge. It is a disassembled perspective view of a cell administration container, a support body, a cartridge, and a changer. It is a perspective view which shows an example of the operation tool which operates a cell administration container. (A), (b) is a front view which shows operation | movement of a changer. (A) is a three-sided view which shows the removal stand which removes a cell administration container, (b) is a three-sided figure which shows an operation jig.

  Hereinafter, the present invention will be described in detail using embodiments shown in the drawings. However, the constituent elements, types, combinations, shapes, relative arrangements, and the like described in this embodiment are not intended to limit the scope of the present invention thereto alone, as long as they are not specifically described, and are merely illustrative examples. .

[Outline of Cell Administration Device]
The cell administration device according to the present invention is a device for seeding cells substantially uniformly in a porous cell support (hereinafter simply referred to as “support”) having a three-dimensional structure.
FIG. 1 is a three-sided view showing the entire cell administration device.
The cell administration device 100 has a support housing recess 22 formed on the upper surface 21 and a slit 23 penetrating the bottom wall of the support housing recess as shown in FIG. A cell administration container 20 for containing the support 10 shown in FIG. 2, a cell suspension feeder 130 for supplying a cell suspension to the support contained in the support containing recess, and the support being impregnated with the support A water absorbent material separating means 190 for bringing a water absorbent material that absorbs water from the induction aqueous solution through the slit into contact with the slit from below.
The cell suspension is a culture solution in which cells to be administered to a support are suspended.

When seeding cells with the support, first, the cell administration container 20 is prepared in which the support 10 impregnated with the induction aqueous solution is accommodated in the support accommodation recess 22. A cell suspension feeder 130 supplies a cell suspension from above the support 10. The water absorbent material separating means 190 brings the water absorbent material into contact with the lower surface of the cell administration container 20 at a predetermined timing, and sucks the induction aqueous solution from the lower surface of the support 10 through the slit 23. By performing the above operation, the cell administration apparatus 100 automatically performs an operation of replacing the cell suspension in the support 10 with the induction solution and seeding the cells substantially uniformly in the support.
The cell administration apparatus and each member involved in cell administration to the support are placed in an isolator in which a sterile environment is maintained by decontamination with hydrogen peroxide vapor or the like, and each operation is performed in the isolator.

  In the following description, the lateral direction of the cell administration apparatus is the width direction (X-axis direction), and the direction orthogonal to this is the depth direction or the longitudinal direction (Y-axis direction). The side in which the worker and the operator face each other is referred to as the front, and the opposite side is referred to as the back.

[Porous cell support]
FIG. 2 is a view showing an example of a porous cell support.
The porous cell support (support) 10 used in the present embodiment is a block having a communicating small pore structure with an average pore diameter of 200 to 300 μm and a porosity of 85 to 91%. The support 10 has an outwardly domed curved surface 11 and a substantially flat surface 12 whose edge is connected to the edge of the curved surface. The flat surface 12 of the support 10 has an oval shape, and the cross-sectional shape of the support 10 when cut along the lateral direction is semicircular.
The size of the support 10 can be, for example, width 6 mm × height 3 mm × length 50 mm. In this case, the volume of the support 10 is about 678 μL, and the volume (the amount of liquid which can be impregnated) in the case where the porosity is 90% is about 610 μL.

[Cell administration container]
FIG. 3 shows an example of a cell administration container.
The cell administration container 20 has a support housing recess 22 formed on the upper surface 21 and a slit 23 penetrating the bottom wall of the support housing recess 22. In the support housing recess 22, the support 10 impregnated with the induction aqueous solution is stored.
The support housing recess 22 has a curved inner surface 24 that is convex downward, and is formed in a shape in which the support 10 fits when the flat surface 12 of the support 10 is placed on top. In other words, the inner surface 24 of the support housing recess 22 has a curved shape that makes surface contact (or close contact) with at least a part of the curved surface 11 when the flat surface 12 is the upper surface and the support 10 is housed. The curved surface 11 of the support 10 and the inner wall of the support housing recess 22 are used in order to allow the cell suspension to penetrate into the support 10 smoothly by using the power of the water-absorbent material to attract the induction aqueous solution. It is preferable to be in close contact. The size of the support housing recess 22 can be set to be 6 mm wide × 8 mm deep × 50 mm long when the support 10 shown in FIG. 1 is housed.

In addition, the slits 23 are formed at positions corresponding to the top of the curved inner surface 24, that is, portions corresponding to the top of the curved surface 11 of the support 10. Therefore, the slit 23 is formed at the lowermost portion of the support housing recess 22 along the longitudinal direction of the support housing recess 22. By forming the slit 23 at the lowermost part of the support housing recess 22, the induced aqueous solution can be absorbed smoothly using attraction. The size of the slit can be set to, for example, 1 mm wide × 44 mm long.
Here, the cell administration container 20 needs to hold the induced aqueous solution impregnated with the support 10 without dropping it from the slit 23 until the water absorbent material is brought into contact. Therefore, it is desirable that the material of the cell administration container 20 be made of a member that has a large contact angle and can be sterilized. For example, if PTFE (polytetrafluoroethylene) or the like is used, the contact angle is large. The inductive aqueous solution impregnated with the support 10 does not leak from the slit and can be sterilized by methods other than gamma ray sterilization such as ethylene oxide gas and hydrogen peroxide gas. The reason for performing sterilization by methods other than gamma ray sterilization is that PTFE is likely to be degraded by gamma ray irradiation and is likely to change its characteristics, but when problems due to changes in characteristics do not occur, for example, the cell administration container should be disposable. If it is done, gamma sterilization may be used. When polyethylene or the like with a small characteristic change of material in gamma ray sterilization is used for a cell administration container, the induction aqueous solution impregnated with the support 10 leaks from the slit 23 and prevents leakage because the contact angle is small. If the width of the slit is narrowed, suction of the inductive aqueous solution is not appropriately performed, which is not preferable as the material of the cell administration container 20.

The depth of the support housing recess 22 is set larger than the height of the support 10. Specifically, the depth of the support housing recess 22 is substantially equal to the volume of the support housing recess 22 plus the volume of the support 10 and the volume of the liquid that can be impregnated by the support 10. Or set to be larger than this. In other words, when the support 10 is accommodated in the support accommodation recess 22, the volume of the support accommodation recess 22 left on the upper side of the support 10 is the volume of the cell suspension that can be absorbed by the support 10. The depth of the support housing recess 22 is set so as to have a volume substantially equal to or larger than the above. By setting the volume of the support housing recess 22 in this manner, the support 10 impregnated with the induction solution is accommodated in the support housing recess 22, and the support 10 is supported on the accommodated support 10. Can be filled with the total amount of cell suspension to be absorbed.
A protrusion 25 projecting downward is formed on the lower surface of the cell administration container 20. The slit 23 is formed in the protrusion 25. A step 26 extending along the slit 23 is formed on the lower surface of the cell administration container 20 by the protrusion 25. By projecting the lower surface of the cell administration container 20 around the slit 23 downward, the water absorbent material can be reliably brought into contact with the slit 23 portion.
At a site of the cell administration container 20 where the support housing recess 22 is avoided, two first positioning holes 27 are formed to penetrate in the vertical direction (the thickness direction of the cell administration container 20). The two first positioning holes 27 are arranged with the support housing recess 22 in between, and are arranged such that the positions of the support housing recess 22 in the longitudinal direction are different. That is, the two first positioning holes 27 are formed in the diagonal direction of the cell administration container 20 substantially rectangular in top view. The first positioning holes 27 prevent erroneous attachment in the vertical direction (upper and lower surfaces) of the cell administration container 20 when the cell administration container 20 is attached to each member.

Further, in the cell administration container 20, two second positioning holes 28 which are different from the first positioning holes 27 are formed penetrating in the vertical direction (the thickness direction of the cell administration container 20). The two second positioning holes 28 are also formed in the diagonal direction of the cell administration container 20 in the same manner as the first positioning holes 27. The second positioning hole is different in diameter from the first positioning hole. The second positioning holes 28 prevent erroneous attachment in the vertical direction of the cell administration container 20 when attaching the cell administration container 20 to each member.
At one corner of the cell administration container 20, a chamfered portion 29 chamfered in a square shape is formed. The chamfered portion 29 prevents erroneous attachment in the front-rear direction of the cell administration container when attaching the cell administration container 20 to each member.
Further, at a portion of the cell administration container 20 where the support housing recess 22 is avoided, the operation rod insertion hole 30 is formed to penetrate along the longitudinal direction of the support housing recess 22. The control rod insertion hole 30 is a hole for inserting a jig for operating the cell administration container 20 without touching the cell administration container 20.

[Container holder]
The container cradle on which the cell administration container is placed will be described. The container pedestal is a member on which the cell administration container is directly mounted, and is used when the cell administration container is installed in the cell administration device. FIG. 4 is a view showing an example of the container holder.
The container holder 40 has a table main body 41 on which the cell administration container 20 is mounted and fixed, and a handle portion 48 connected to the table main body 41, and is formed of a metal material such as stainless steel.

<Base body>
The table main body 41 has a substantially rectangular plate shape, and holds the cell administration container 20 without interfering with the communication state of the slit 23 of the cell administration container 20 in the vertical direction. That is, a rectangular through hole 42 which exposes the slit 23 to the lower side of the table body 41 is formed in the in-plane central portion of the table body 41. The through hole 42 is formed in a size in which the protrusion 25 of the cell administration container 20 can be inserted. The thickness of the table main body 41 is set to be substantially equal to or thinner than the amount of protrusion of the protrusion 25, and the slit 23 protrudes at a position substantially equal to or lower than the lower surface of the table main body 41 Do.
On the upper surface of the table main body 41, a position determination pin 43 for determining the position and the upper and lower direction of the cell administration container 20 and a direction determination pin 44 for determining the mounting direction (front and rear direction) of the cell administration container 20 are protruded. . The position determination pin 43 is inserted into the first position determination hole 27 of the cell administration container 20 to determine the position and the top and bottom orientation of the cell administration container 20. In addition, the direction determining pin 44 has the mounting direction of the cell administration container 20 normally mounted at the position and the top and bottom direction of the cell administration container 20 where the position determination pin 43 can be inserted into the first positioning hole 27 In this case, it does not interfere with the chamfered portion 29 of the cell administration container 20, and is formed at a position where it interferes with corner portions other than the chamfered portion 29 when the mounting direction is incorrect.

At the rear end edge of the table main body 41, when the container receiving table 40 is installed in the cell administration device, position determination to define the mounting position of the table main body 41 in the depth direction with respect to the cell administration device A recess 45 is formed. The position determination recess 45 is formed in two places separated in the width direction. By providing a plurality of positioning recesses 45, the mounting angle of the table body 41 can be easily defined.
A deep portion 45a of the positioning recess 45 has a shape in which a positioning protrusion 116 provided in the cell administration device 100 is in close contact, and prevents rattling of the table body 41 with respect to the cell administration device. Further, a guide portion 45b whose width gradually decreases toward the back portion 45a is formed on the opening end side of the position determination recess 45, and the position determination protrusion 116 is smoothly guided toward the back portion 45a.

<Handle part>
The handle portion 48 projects forward from the front end edge of the base body 41, that is, the edge opposite to the edge where the positioning recess 45 is formed. The handle portion 48 is continuously provided in a stepped state with respect to the table main body 41 so as to be positioned above the upper surface of the table main body 41. When the container pedestal 40 is placed on the workbench, the lower surface of the stand main body 41 is in close contact with the workbench, while the handle portion 48 is separated from the workbench and floats up. Therefore, the handle portion 48 can be easily grasped by the glove installed in the isolator or a dedicated grasping tool, and the operation of the container pedestal 40 can be facilitated.

[Stabilizer]
The stabilizer placed on the cell administration container 20 fixed to the container holder 40 will be described. FIG. 5 is a view showing an example of the stabilizer.
When the water absorbing material separating means 190 brings the water absorbing material into contact with the slit 23 from below with the stabilizer 80, the pressing force from the water absorbing material separating means 190 causes the cell administration container 20 to be lifted from the container stand 40. It is a weight to prevent it and is made of a metal material such as stainless steel.
The stabilizer 80 has a through hole 81 for exposing the support housing recess 22 of the cell administration container 20 upward, and a position determination hole 82 through which the position determination pin 43 of the container holder 40 is inserted. It has substantially the same shape as the administration container 20. The lower surface of the stabilizer 80 in contact with the upper surface 21 of the cell administration container 20 is a flat surface. A recessed line 83 extending along the depth direction is formed on the top surface of the stabilizer 80 including the formation site of the through holes 81 and in the central portion in the width direction. The concaves 83 are formed to prevent interference with the cell suspension feeder 130 when administering cells to the support 10.

〔Syringe〕
The syringe will be described. FIG. 6 is a front view of a syringe.
The syringe 50 is installed in the cell administration device in a state in which the cell suspension is filled, and is used when the administration cell and the culture solution are mixed to produce a cell suspension.
The syringe 50 has an outer cylinder 51 and a piston 55.
The outer cylinder 51 includes an outer cylinder main body 52 formed of a hollow cylindrical body in which a cell suspension is filled, and a nozzle disposed at the tip of the outer cylinder main body 52 and discharging the cell suspension in the outer cylinder main body 52 A portion 53 and a finger flange 54 protruding in the outer diameter direction of the outer cylinder main body 52 at the base end of the outer cylinder main body 52 are provided.
The piston 55 is provided with a gasket 56 that moves back and forth in the outer cylinder main body 52 in close contact with the inner surface of the outer cylinder main body 52 at the tip end portion, and has a piston head 57 for pressing the piston 55 at the base end. When the piston 55 advances toward the nozzle portion 53 along the axial direction of the outer cylinder 51, the contents (administered cells, culture fluid, or cell suspension) filled in the outer cylinder main body 52 are pushed out. The nozzle portion 53 discharges the ink to the outside. Further, when the piston 55 retracts from the nozzle portion 53, the liquid is introduced into the outer cylinder main body 52 through the nozzle portion 53.

[Cell mixer]
A cell mixer will be described which uniformly mixes the administered cells and the culture solution to produce a cell suspension. FIG. 7 is a perspective view showing an example of a cell mixer. In the following description, the axial direction of the syringe is described as the axial direction of the cell mixer.
The cell mixer 60 repeatedly moves the outer cylinder 51 of the two syringes 50a and 50b connected in a state in which the inside of the hollow portion is communicated by the three-way stopcock 61 with respect to the piston 55 for a predetermined time repeatedly. , 50b are uniformly mixed with the administration cells and the culture solution.
The first syringe 50a is filled with a prescribed amount (for example, 1.5 ml) of administered cells, and the second syringe 50b is filled with a prescribed amount (for example, 3.0 ml) of culture solution. The nozzle portions 53 of the two syringes 50a and 50b are connected by the three-way stopcock 61 to form a syringe unit 62 in which the hollow portions of the two syringes communicate with each other.

The cell mixer 60 is disposed on the housing 63 and the upper surface of the housing 63 to fix the positions of the piston heads 57 of the respective syringes 50a and 50b constituting the syringe unit 62. , 64b and the first and second head support portions 64a and 64b, so as to axially reciprocate the outer cylinder 51 and the three-way stopcock 61 of the first and second syringes 50a and 50b. And a slide table 67 for supporting.
Each head support portion 64 a, 64 b includes a head holding member 65 for holding each piston head 57, and the head holding member 65 is configured to be able to advance and retract in the axial direction by a screw member 66. Thus, the position of the head holding member 65 can be finely adjusted corresponding to the position of the piston head 57 which changes in accordance with the amount of liquid filled in each of the syringes 50a and 50b.
The slide table 67 supports the stopper support recess 68 supporting a portion of the three-way stopcock 61 projecting in the direction intersecting the axial direction of the syringe 50 and the axial direction supporting the distal end side of the outer cylinder 51 of each of the syringes 50a and 50b. The first and second outer cylinder support recesses 69a, 69b are extended. The outer cylinder 51 and the three-way stopcock 61 of each of the syringes 50 a and 50 b are fixed to the slide table 67.

Further, the slide base 67 is axially reciprocated by an air cylinder which is a drive means housed in the housing 63. The air that is the driving source of the air cylinder is a synthetic air mixed with approximately 21% oxygen and approximately 79% nitrogen of the Japanese Pharmacopoeia, and prevents contamination of each part when gas leaks due to damage to the air cylinder. .
The slide table 67 is controlled by the control device to reciprocate at a predetermined time interval (for example, one reciprocation in two seconds) for a predetermined time (for example, five minutes). As the slide table 67 reciprocates, the administration cell and the culture solution reciprocate the two syringes 50a and 50b, and in the process, the administration cell and the culture solution are uniformly mixed. All cell suspensions prepared are collected into one syringe, which is placed in the cell administration device.
In this cell mixer 60, the position of the piston head 57 at the axial end of the syringe unit 62 is fixed, and the outer cylinder 51 is moved relative to the piston 55. Therefore, the total axial length of the cell mixer 60 Can be shortened. Thus, no extra space is taken in the isolator, which has a limited working space. In addition, since the axial intermediate portion of the syringe unit 62 is moved, the number of parts for transmitting the driving force is one, so that the configuration of the apparatus can be simplified.

[Cell administration device]
The cell administration device will be described with reference to FIG.
The cell administration apparatus 100 includes a cell administration container setting unit 110 for placing the cell administration container 20, a cell suspension feeder 130 for supplying a cell suspension to a support housed in the cell administration container 20, and a cell suspension unit 130. Cell suspension feeder drive means (feeder drive means 160) for driving the suspension feeder 130 in the X-axis direction, Y-axis direction and Z-axis direction, and the water absorbing material from the lower side of the cell administration container And a water absorbent material separating means 190.
The cell administration container setting unit 110, the feeder driving unit 160, and the water absorbing material contacting and separating unit 190 are fixed in position on the base plate 101. In addition, the cell suspension feeder 130 is supported by the feeder driving means 160.

<Cell administration container setting part>
FIG. 8 is a three-sided view showing the cell administration container setting unit and the water absorbent material contacting / separating means. As shown in FIG. 1, the cell administration container placement unit 110 is a site that holds the container pedestal 40 on which the cell administration container 20 is mounted. In addition, the container holder 40 is abbreviate | omitted in the top view and right view of FIG.
The cell administration container setting unit 110 includes a base 111 whose longitudinal direction is the width direction, and the base 111 is supported by a plurality of leg members 112. The leg members 112 are fixed to the base plate 101.
The base 111 is provided with a plurality of slots 113 (five slots in the figure) along the width direction, each of which comprises a recess 113 into which the container holder 40 can be inserted and removed freely. Each slot 113 has an insertion opening 114 for inserting the container holder 40 at the front end.

The slot 113 includes guide rails 115 extending in the depth direction at both ends in the width direction. At the deepest portion of the slot 113, a positioning protrusion 116 for determining the position of the container holder 40 in the depth direction is formed to project upward from the base 111. The slit 23 of the cell administration container 20 mounted in the slot 113 via the container holder 40 faces the water absorbent material separating means 190 located below the base 111.
The guide rail 115 slidably guides the container support 40 in the depth direction by sandwiching the width direction end of the container support 40 in the vertical direction.
The positioning protrusion 116 closely contacts the back 45 a of the positioning recess 45 of the container pedestal 40 to thereby determine the insertion position of the container pedestal 40 in the depth direction with respect to the slot 113. Further, the guide portion 45b of the positioning recess 45 of the container holder 40 is formed wider than the diameter of the positioning protrusion 116, and the width gradually decreases toward the back portion 45a. Even when inserted obliquely into the slot 113, the positioning protrusion 116 is smoothly guided toward the back portion 45a.
The guide rails 115 and the positioning projections 116 cooperate to prevent the container cradle 40 from tilting.

Cell suspension feeder
FIG. 9 is a trihedral view showing a cell suspension feeder.
The cell suspension feeder 130 is a device for feeding the cell suspension filled in the syringe 50 to the support 10.
The cell suspension feeder 130 is disposed at the lower part of the outer surface of the housing 131 facing the worker, and the syringe holder 135 for holding the syringe 50, and the piston of the syringe 50 held by the syringe holder 135. And a head pressing mechanism 151 for pressing the head 57. A guide elongated hole 133 is vertically formed in the front panel 132 of the housing 131 in the vertical direction, and guides the head pressing member 156 constituting the head pressing mechanism 151 in the vertical direction.

Syringe holder
The syringe holder 135 includes an upper holder 136 for holding the upper portion of the outer cylinder 51 of the syringe 50 (the outer cylinder main body 52 located on the finger flange 54 side) and a lower holder 141 for holding the lower portion of the outer cylinder 51 (nozzle portion 53). And hold the syringe 50 with the nozzle 53 directed downward.

The upper holder 136 and the lower holder 141 respectively include two arm members 137 and 142 that project in parallel at a predetermined distance from the front panel 132 of the housing 131. The distance between the arm members 137 is set to be substantially the same as or slightly larger than the diameter of the outer cylinder main body 52. Further, the distance between the arm members 142 is set to be substantially the same as or slightly larger than the diameter of the nozzle portion 53. The two arm members 137 and 142 hold the syringe 50 by sandwiching the outer cylinder 51 (the outer cylinder main body 52 and the nozzle portion 53).
At the tip of the arm members 137 and 142 and on the surface facing the outer cylinder 51, stoppers 138 and 143 made of metal balls configured to be able to move in and out are disposed. The stoppers 138 and 143 are elastically urged in the projecting direction, that is, the direction in which the outer cylinder 51 is pressed by an elastic urging member such as a spring disposed inside the arm members 137 and 142.
When mounting the syringe 50 in each syringe holder 135, the outer cylinder 51 is inserted between the two arm members 137 and 142 in a state where the syringe 50 is erected with the nozzle portion 53 facing downward. The stoppers 138 and 143 are pushed by the outer surface of the outer cylinder 51 and inserted into the arm members 137 and 142 in the process of inserting the syringe 50, and protrude from the arm members 137 and 142 when the syringe 50 is held. Prevent clatter and falling off.

The upper holder 136 functions as a means for positioning the syringe 50 in the vertical direction. That is, by bringing the lower surface of the finger flange 54 into close contact with the upper surface of the upper holder 136 at the time of mounting the syringe 50, the vertical position of the syringe 50 with respect to the syringe holder 135 is determined.
Here, the amount of the cell suspension to be discharged with respect to the axial movement distance of the piston 55 is determined by the diameter (inner diameter and outer diameter) of the outer cylinder main body 52 of the syringe 50. The syringe holder 135 is designed to be capable of holding only a single type of syringe 50 (for example, only a syringe for 5 ml) in order to prevent operation errors such as excessive or insufficient amount of cell suspension supplied to the support.

<< Head pressing mechanism >>
The head pressing mechanism 151 pressing the piston head 57 has a drive motor 153 housed in the lower part in the housing 131 so that the output shaft 152 is in the vertical direction, and a relative rotation coaxial with the output shaft 152 of the drive motor 153 And a head pressing member 156 fixed to the female screw member 155 to press the upper surface of the piston head 57. The head pressing mechanism 151 drives the head pressing member 156 by a so-called feed screw method. Further, an adjusting head 157 attached to the upper end portion of the screw rod 154 so as not to be relatively rotatable with respect to the screw rod 154 and exposed from the top surface of the housing 131 is provided.

The head pressing member 156 is connected to a female screw member 155 disposed inside the housing 131 via a vertically extending guide long hole 133 formed through the front panel 132 of the housing 131, and the head pressing member The moving direction of the screw 156 and the female screw member 155 is restricted by the guide elongated hole 133 in the vertical direction.
The drive motor 153 is, for example, a five-phase stepping motor. The screw rod is formed with a screw thread so as to move the female screw member 155 in the axial direction by a predetermined distance (for example, 1 mm) every one rotation. The control means for controlling the drive motor 153 controls the moving distance of the female screw member 155 and the head pressing member 156 based on the rotation angle of the output shaft 152 of the drive motor 153.
The adjusting head 157 is a cylindrical member having an outer peripheral surface subjected to anti-slip processing such as knurling. The adjusting head 157 is set to a size that allows the operator to easily rotate the adjusting head 157 with the glove installed in the isolator. The adjustment head 157 can freely rotate when no current flows in the drive motor 153. The adjustment head 157 is used to manually finely adjust the position of the head pressing member 156 with respect to the piston head 57 of the syringe 50 held by the syringe holder 135.
When the drive motor 153 rotates, the screw rod 154 rotates in conjunction with the rotation of the output shaft 152. The female screw member 155 screwed into the screw rod 154 moves in the axial direction of the screw rod 154 without rotating because the moving direction is restricted in the vertical direction by the guide elongated hole 133. The head pressing member 156 moves up and down together with the female screw member 155. When the head pressing member 156 is lowered together with the female screw member 155, the piston head 57 is pressed to push the cell suspension filled in the syringe 50 to the outside. From the syringe 50, a cell suspension of an amount according to the movement distance of the piston head 57 pressed by the head pressing member 156 is discharged.

<Cell suspension feeder drive means>
The cell suspension feeder drive means (feeder drive means) will be described with reference to FIG. Hereinafter, the left and right direction is taken as the X axis, the depth direction as the Y axis, and the vertical direction as the Z axis when viewed from the worker (the front of the isolator).
The feeder driving unit 160 includes an X-axis stage 161x, a Y-axis stage 161y, and a Z-axis stage 161z, which move the cell suspension feeder 130 in the X-axis direction, the Y-axis direction, and the Z-axis direction, respectively. There is.
The stages 161x, 161y, and 161z have the same configuration, and the housings 162x, 162y, and 162z, which extend in the axial direction, and the sliders 163x, 163y, and 163z, which support the support and move back and forth along the axial direction. Is equipped. In each housing 162x, 162y, 162z, a stage drive motor, a feed screw rotationally driven by the stage drive motor, a female screw member screwed with the feed screw, and a guide for guiding the sliders 163x, 163y, 163z The rails are built in, and the sliders 163x, 163y and 163z advance and retract in the respective axial directions as the feed screw rotates forward and reverse. Further, the positions of the sliders 163x, 163y, and 163z in the respective axial directions are controlled by the control unit based on the rotation angle of the output shaft of the stage drive motor.

The connection relationship of each member is as follows. The slider 163x of the X-axis stage 161x supports a suitable position of the housing 162x of the Y-axis stage 161y as a supported object, and the Y-axis stage 161y advances and retracts in the X-axis direction. The slider 163y of the Y-axis stage 161y supports an appropriate position of the housing 162x of the Z-axis stage 161z as a supported object, and the Z-axis stage 161z advances and retracts in the Y-axis direction. The slider 163z of the Z-axis stage 161z supports the appropriate position of the housing 131 of the cell suspension feeder 130 as a support, and the cell suspension feeder 130 advances and retracts in the Z-axis direction.
With the above configuration, the cell suspension feeder 130 can freely move in the three axial directions of the X axis, the Y axis, and the Z axis.

<Water-absorbent material / Water-absorbent material contact / separation means>
A water absorbing material contacting / separating means for bringing a water absorbing material absorbing water through the slit into contact with the slit from the lower side with respect to the slit and a means for holding the water absorbing material will be described.

<< Water-absorbent material / water-absorbent material holding member >>
FIG. 10 is a view showing a water absorbing material holding member, (a) is a three-sided view showing a water absorbing material holding base, (b) is a plan view showing a pressing frame, (c) holds a water absorbing material It is a cross-sectional view of the water absorbent material holding member.
For example, filter paper can be used as the water absorbing material P that absorbs water in the induction aqueous solution. The water absorbing material P can be used without particular limitation as long as it is a material capable of absorbing the induction aqueous solution as appropriate from the support. The water absorbent material P is mounted on the water absorbent material separating means 190 in a state of being held by the water absorbent material holding member 170.

As shown in FIG. 10, the water absorbent material holding member 170 holding the water absorbent material P is a water absorbent material holding base 171 on which the water absorbent material P is placed and a schematic rectangular frame in plan view mounted on the water absorbent material holding base 171. And a holding frame 176 in the shape of a circle.
The water-absorbent material holding base 171 is provided with a thick-walled central portion 172 with a top surface projecting from the central portion 172 and a thin outer peripheral portion 173 with respect to the central portion 172, and a water-absorbent material holding portion 174 substantially rectangular in plan view A grip handle 175 is provided continuously with one end of the material holding portion 174 in the longitudinal direction. The upper surface of the central portion 172 is a flat surface.
The pressing frame 176 has an opening 177 for exposing the central portion 172 of the water absorbent holding base 171 at the central portion, and is superimposed on the outer peripheral portion 173 of the water absorbent holding base 171. The pressing frame 176 sandwiches the water absorbing material P with the outer peripheral portion 173 to prevent the water absorbing material P from shifting or floating, and maintains the surface of the water absorbing material P in contact with the slit in a planar shape.
The handle 175 is a portion for holding when operating the water absorbent material holding member 170, such as when the water absorbent material holding member 170 is installed on the water absorbent material contacting / separating means 190.

<< Cushioning material >>
When the water absorbing material holding member 170 is attached to the water absorbing material contacting means 190, the cushion material 180 is disposed between the water absorbing material holding member 170 and the upper plate 192 of the water absorbing material contacting means 190.
FIG. 11 is a view showing an example of the cushioning material.
The cushioning material 180 plays a role of bringing the water absorbing material P into close contact with the projecting portion 25 of the cell administration container 20 by elastic force when the water absorbing material P absorbs the induced aqueous solution.
The cushion material 180 is a sheet material made of an elastic material such as silicon sponge, for example, and the planar shape is substantially the same as the water absorbent material holding member 170. At the front end portion and the rear end portion of the upper surface of the cushion material 180, protrusions 181 extending in the width direction are formed.
The contact area between the cushion material 180 and the water absorbent material holding member 170 can be reduced by the ridges 181. By reducing the contact area, the load per unit area applied to the contact portion with the water absorbent material holding member 170 is increased, and the deformation amount of the cushion material 180 is secured. Thereby, non-uniform adhesion (uneven pressing force) of the water-absorbent material P with respect to the protrusion 25, wrinkles of the water-absorbent material, accuracy errors of the respective members, and the like are absorbed, and non-uniform absorption of the induced aqueous solution is suppressed.

"Water-absorbent material separation means"
The water absorbent material separating means 190 will be described with reference to FIG.
The water absorbent material separating means 190 is provided below the base 111 constituting the cell administration container setting unit 110 for each slot 113 corresponding to each cell administration container 20.
Each water absorbing material contacting means 190 comprises a water absorbing material set base 191 provided with an upper plate 192 on which the water absorbing material holding member 170 holding the water absorbing material P is placed, and the water absorbing material P in the slit 23 of the cell administration container 20. And an air cylinder 205 for displacing the water absorbent material set base 191 between an approach posture for bringing the water absorbent material into close contact and a separation posture for bringing the water absorbent material away from the slit.

Water-absorbent material set base
The water absorbent material set base 191 includes an upper plate 192 on which the water absorbent material holding member 170 is mounted, and two side plates 193 continuously provided at respective end portions in the width direction of the upper plate 192 and hanging downward.
At a front end portion and a rear end portion of the upper plate 192, a front position determination pin 194 and a rear position determination pin 195 for determining the positions of the cushion material 180 and the water absorbent material holding member 170 in the front and rear direction project upward. The front position determining pin 194 and the rear position determining pin 195 are arranged two by two in the width direction, respectively.
A notch 196 is formed at the center in the width direction of the front end edge of the upper plate 192. The deepest portion of the notch 196 is located rearward of the front position determination pin 194, and interferes with the upper plate 192 when the cushioning material 180 is held by tweezers or the like and placed on the upper plate 192. To prevent.
A guide piece 197 for determining the position in the width direction of the cushion material 180 and the water absorbent material holding member 170 is attached to a middle portion in the depth direction of each side plate 193 so as to protrude above the upper plate 192. The upward projecting amount of the guide piece 197 is set so as not to contact the container pedestal 40 inserted in the slot 113 when the water absorbent material set base 191 is displaced to the approach posture.
The four positioning pins 194 and 195 provided on the upper plate 192 and the two guide pieces 197 determine the positions of the cushion 180 and the edge of the water absorbent material holding member 170, and both are positioned on the upper plate 192.

Each side plate 193 of the water absorbent material set base 191 is rotatably supported by a pivot shaft member 201 which is supported by the base 111 and extends substantially horizontally in the width direction at a rear end proper position. The water absorbent material set base 191 is supported so that the front end moves up and down around the pivot shaft member 201.
The water absorbent material set base 191 is disposed below the pivot shaft member 201, and includes a pressed member 202 to which the driving force (pressing force) from the air cylinder 205 which is the driving means is transmitted. In this example, the pressed member 202 is a shaft member that extends substantially horizontally in the width direction and connects the two side plates 193. When the pressed member 202 is pressed by the air cylinder 205, the water absorbent material set base 191 pivots upward about the pivot shaft member 201, and the upper plate 192 ascends. The upper surface of the upper plate 192 is substantially parallel (substantially horizontal) to the lower surface (projecting portion) of the cell administration container 20 in the approaching posture, and in the separated posture, the front end is lowered and inclined.
If it is attempted to raise the upper surface of the upper plate 192 linearly while keeping the horizontal posture straight, the water absorbent material P may contact the lower surface of the cell administration container 20 in an inclined state to cause water absorption unevenness of the induction aqueous solution. There is. In the present embodiment, since the upper plate 192 is raised toward the lower surface of the cell administration container 20 while rotating the water absorbent material set base 191, the water absorbent material when the water absorbent material P contacts the lower surface of the cell administration container 20 It prevents inclination of P and effectively prevents uneven water absorption of the induction aqueous solution.

<< drive means >>
The driving means for moving the water absorbent material set base up and down is the air cylinder 205 in this embodiment. The air cylinder 205 includes a cylinder tube 206 into which driving air is introduced, and a piston rod 207 which protrudes from and retracts from the cylinder tube 206, and is fixed to the base plate 101. The mounting position of the air cylinder 205 and the elevation angle with respect to the base plate 101 are set such that the direction of the piston rod 207 is substantially tangent to the arc-like locus drawn by the pressed member 202 about the pivot shaft member 201. Ru.
When the piston rod 207 protrudes, it presses the pressed member 202 to displace the water absorbent material set base 191 in the approaching posture. Also, when the piston rod 207 retracts, the water absorbent material set base 191 is displaced to the retracted position by its own weight.
The cylinder tube 206 is filled with synthetic air mixed with about 21% oxygen and about 79% nitrogen of the Japanese Pharmacopoeia. By using synthetic air as a gas for driving the air cylinder 205, the contamination of each part when the gas leaks into the isolator due to the damage of the air cylinder 205 is prevented.

<Control device>
A control device that controls the operation of each unit of the cell administration device is configured of a computer device provided with a central processing unit (CPU), a read only memory (ROM), a random access memory (RAM), and the like. A control program for controlling each part of the cell administration device is stored in the ROM. The programs stored in the ROM are booted into the RAM, and when the programs are started, the RAM becomes a work area by the CPU of these programs. The CPU boots the program in the ROM to the RAM, and executes each program while using the RAM as a work memory.

[Operation of Cell Administration Device]
The preparation for performing the cell administration operation to the support 10 by the cell administration device 100 is as follows.
First, the cushioning material 180 is placed on the upper plate 192 of the water absorbent material set base 191. The water absorbent material P is sandwiched between the water absorbent material holding base 171 and the pressing frame 176, and the water absorbent material holding member 170 in a state of holding the water absorbent material P is placed on the cushion material 180.
The support 10 immersed in the induction aqueous solution is taken out by tweezers or the like, and the flat surface 12 is accommodated in the support accommodating recess 22 as the upper surface. The support 10 accommodated in the support accommodation recess 22 is pressed by the plunger 320 functioning as a positioning jig as shown in FIG. 13 so that the flat surface 12 is substantially horizontal in the support accommodation recess 22. And so that the support 10 is disposed at a predetermined depth position of the support housing recess 22. The plunger 320 has a flat surface 321 having substantially the same shape as the flat surface 12 of the support 10.
The cell administration container 20 in which the support 10 is accommodated in the support accommodation recess 22 is placed and fixed on the container pedestal 40. That is, the position determination pin 43 is inserted into the first position determination hole 27 in a state where the position determination pin 44 of the container holder 40 and the position of the chamfered portion 29 of the cell administration container 20 are aligned. The container holder 40 is inserted into the slot 113 by holding the handle portion 48 of the container holder 40 on which the cell administration container 20 is mounted and fixed indirectly with a device or directly with a glove. At this time, the container support 40 is inserted along the guide rail 115 such that each widthwise end of the container support 40 is sandwiched by the guide rails 115. The positioning protrusion 116 of the slot 113 is brought into close contact with the positioning recess 45 of the container pedestal 40 to determine the position of the cell administration container 20 within the slot 113.

When the manual set switch for driving the cell suspension feeder driving means 160 is pressed, the control means drives the feeder driving means 160 to operate the cell suspension feeder 130 from the standby position shown in FIG. Move to the manual setting position on the side (Y-axis direction). The manual setting position is a position where the cell suspension feeder 130 can be operated using the glove of the isolator. When moving the cell suspension feeder driving means 160 to the manual setting position, the control means controls the driving motor 153 so as to raise the head pressing member 156. Below the head pressing member 156, a gap to which the syringe 50 filled with the cell suspension can be attached is formed.
The syringe 50 filled with the cell suspension is attached to the syringe holder 135 of the cell suspension feeder 130 at the manual setting position. At the time of mounting, the lower surface of the finger flange 54 is brought into close contact with the upper surface of the upper holder 136 to determine the vertical position of the syringe 50 with respect to the syringe holder 135. After mounting, the adjustment head 157 is rotated to bring the head pressing member 156 into close contact with the piston head 57. At this time, the position of the head pressing member 156 is adjusted to such an extent that the cell suspension is slightly exposed to the outside from the nozzle portion 53.
Finally, when the set switch is pressed, the control means drives the feeder driving means 160 to move the cell suspension feeder 130 to the standby position.

The cell administration operation to the support 10 by the cell administration device 100 is as follows. The following operation is performed by the control unit drivingly controlling the feeder drive unit 160 and the drive motor 153 of the head pressing mechanism 151. Also, the portion of the syringe 50 held by the cell suspension feeder 130 moves along the movement trajectory shown in FIG.
When the start switch is pressed, the cell suspension feeder 130 advances from the standby position. At this time, the cell suspension feeder 130 moves on a position where the nozzle portion 53 does not face the support housing recess 22, for example, on the guide rail 115. When the cell suspension feeder 130 reaches a position corresponding to the front end of the support housing recess 22, the cell suspension feeder 130 has the nozzle 53 on the front end of the support housing recess 22. Move to the left in the figure to be positioned. The cell suspension feeder 130 is lowered, and the nozzle portion 53 is positioned directly above the support 10 in the support housing recess 22. Cell suspension feeder 130 retracts at a predetermined rate.
At this time, the head pressing mechanism 151 presses the piston head 57 at a predetermined speed so that the cell suspension of an amount according to the retraction speed of the cell suspension feeder 130 is discharged from the nozzle unit 53.
When the nozzle portion 53 reaches the rear end of the support housing recess 22, the cell suspension feeder 130 is raised. The cell suspension feeder 130 is moved onto the guide rail 115 of the adjacent slot 113 on the left side in the figure.
While repeating the above operation, the cell suspension feeder 130 moves across the plurality of cell administration containers 20 to administer the cell suspension to the support 10 in each cell administration container 20. Do. In addition, when the administration of the cell suspension to all the supports 10 is finished, the cell suspension feeder 130 rises and moves to the standby position.

The water absorbent material separating means 190 raises the water absorbent material P toward the slit 23 of the cell administration container 20 containing the support 10 at the timing when the administration of the cell suspension to the certain support 10 is finished. The water absorbing material P is brought into contact with the slit 23 about 0.7 seconds (at least within 1.0 second) after the end of the administration of the cell suspension. The time during which the water absorbing material P is in contact with the slit 23 is approximately 1.8 seconds to 3.2 seconds. Since the amount of liquid absorbed by the water absorbing material P is substantially proportional to the time of contact with the slit 23, the amount of water absorbed by the water absorbing material P is the time during which the water absorbing material P is in contact with the slit 23 Control based on In addition, since it is difficult to absorb water only in the induction aqueous solution, the time for which the water absorbing material P is in contact with the slit 23 should be set so that all the induction aqueous solution and a part of the cell suspension absorb some water. Is desirable.
The water absorption of the induction aqueous solution starts from the administration of the cell suspension to the start of the fall of the induction aqueous solution by gravity. In addition, since there is a variation in the number of cells administered to the support 10 if there is a variation in the time from the administration of the cell suspension to the absorption of water in the induction aqueous solution, the driving of the water absorbing material contacting means 190 is supported. It is carried out every body 10.

〔Example〕
Hereinafter, the cell administration container 20 shown in FIG. 3 (size of support accommodation recess: width 6 mm × depth 8 mm × length 50 mm, slit size: width 1 mm × length 44 mm) and the cell administration device 100 shown in FIG. The results of administering the cells to the support 10 (width 6 mm × height 3 mm × length 50 mm, porosity 87%) shown in FIG.

When the number of living cells contained in the support was measured, it was found that the variation between the supports was as small as 2.9% in CV value, and the cells were uniformly administered to the support.
The first support is the support housed in the cell administration container 20 placed at the right end in FIG. 1, and the fifth support is placed in the cell administration container 20 placed at the left end in FIG. It refers to the housed support.

〔effect〕
It is necessary to produce a plurality of (for example, 5) supports for each patient, including those for which the cells have been administered, including those for preparation and for examination. However, it is necessary to prevent quality variations from occurring among these products.
According to this embodiment, by mechanizing the complicated cell administration operation, the product can be homogenized as compared with the case of manufacturing the support by human operation. In addition, since production of a support on which cells are seeded can be mechanized, a large amount of support can be produced in a short time.
In the above description, the support impregnated with the induction aqueous solution is accommodated in the support accommodation recess of the cell administration container, but the support before impregnation of the induction aqueous solution is accommodated in the support accommodation recess, and The support may be impregnated with the induction aqueous solution by placing and fixing the administration container on the container pedestal and immersing the container pedestal together with the induction aqueous solution.

〔cartridge〕
The following describes a cartridge for carrying a support while carrying out sterilization treatment with gamma rays or the like in a state of containing a support not impregnated with an induced aqueous solution and maintaining the sterilization state.
FIG. 12 is a perspective view of the cartridge. FIG. 13 is an exploded perspective view of a cell administration container, a support, a cartridge and a changer. In the following description, the longitudinal direction, the latitudinal direction, and the thickness direction are defined based on the shape of the support housed in the cartridge.
The cartridge 300 has a case 310 in which a support housing hole 311 for housing the support 10 is formed penetrating in the thickness direction, and a state in which the support 310 is fitted into the support housing hole 311 from one surface side (lower surface side) of the case 310. A plunger 320 to be inserted, a cover member 330 for covering the support housing hole 311 on the other surface side (upper surface side) of the case 310, and a fixing pin 340 for fixing the cover member 330 to the case 310 are provided.

<Case>
The case 310 has a rectangular plate shape in plan view, and has a shape similar to that of the cell administration container 20 shown in FIG.
The case 310 has an oblong support housing hole 311 formed through in the vertical direction (thickness direction), and a first positioning hole 312 formed in the vertical direction in a portion avoiding the support housing hole 311. And a second positioning hole 313. The support housing hole 311 is formed to extend along the longitudinal direction of the case 310. In addition, the case 310 includes a recess 314 formed on one end surface in the longitudinal direction, and a pin hole 315 which is penetratingly formed in the vertical direction (thickness direction) at the other end in the longitudinal direction and into which the fixing pin 340 is inserted.
The support housing hole 311 has a shape that matches the shape of the flat surface 12 of the support 10, and the size of the case 310 in the thickness direction is constant.

The first positioning hole 312 and the second positioning hole 313 are respectively the same size as the first positioning hole 27 and the second positioning hole 28 formed in the cell administration container 20, The positional relationship of the positioning holes is the same between the case 310 and the cell administration container 20. Therefore, when the case 310 is superimposed on the cell administration container 20, the first positioning holes 27, 312 and the second positioning holes 28, 313 communicate with each other. Further, when the case 310 is superimposed on the cell administration container 20 in a state of being positioned by each positioning hole, the support accommodation hole 311 communicates with the support accommodation recess 22 of the cell administration container 20.
The recess 314 determines the mounting position of the cover member 330 with respect to the case 310.
The fixing pin 340 is inserted into the pin hole 315 in communication with the pin hole 336 formed in the cover member 330.

<Plunger>
The plunger 320 has a substantially elongated cylindrical shape with the flat surface 321 having the same shape as the flat surface 12 of the support 10 as the upper and lower surfaces. Guide grooves 322 are formed along the longitudinal direction on both side surfaces of the middle portion of the plunger 320 in the thickness direction (vertical direction). The plunger 320 is symmetrical in the thickness direction about the guide groove 322. When the case 310, the plunger 320, the cover member 330, and the fixing pin 340 are assembled as the cartridge 300 from the flat surface 321 to the guide groove 322, the insertion is performed while being fitted in the support housing hole 311 It is the part 323.
Although the total thickness of the plunger 320 is thicker than the case 310, the insertion part 323 is thinner than the case 310. In a state in which only the insertion portion 323 is inserted into the support housing hole 311, a space for housing the support 10 is formed by the flat surface 321 located in the support housing hole 311 and the inner side surface of the support housing hole 311. Ru. The plunger 320 supports the flat surface 12 of the support 10 from below.

<Cover member>
The cover member 330 is a member that closes the support housing hole 311 into which the plunger 320 is inserted and prohibits the plunger 320 from moving up and down.
The cover member 330 is adjoined to the upper surface (other surface) of the case 310 to close the support housing hole 311 and is attached to the lower surface (one surface) of the case 310 and exposed from the lower surface of the case 310 The fixed piece 332 provided with the guide slit 333 sandwiching the guide groove 322 portion of the plunger 320, and the connection piece 334 which connects the lid 331 and the fixed piece 332 at one end in the longitudinal direction and is fitted to the recess 314 of the case 310. And an operation piece 335 which extends from one end of the fixed piece 332 over the connection piece 334.

The width of the guide slit 333 is substantially equal to the width of the guide groove 322. Further, the thickness of the fixing piece 332 is substantially equal to the length in the thickness direction of the guide groove 322. The cover member 330 slides along the longitudinal direction of the case 310 by the fixed piece 332 being guided along the longitudinal direction by the guide groove 322 of the plunger 320.
The lid 331 and the fixing piece 332 are attached to the upper surface of the case 310 where the first positioning hole 312 and the second positioning hole 313 are avoided. That is, the width (length in the lateral direction) of the lid 331 and the fixing piece 332 is shorter than the lateral length of the case 310 and interferes with both the first positioning hole 312 and the second positioning hole 313. Not formed in size.
The lid 331 is provided with a pin hole 336 communicating with the pin hole 315 of the case 310 at the other end in the longitudinal direction when the connection piece 334 is fitted in the recess 314. By inserting the fixing pin 340 into both the pin holes 315 and 336, the cover member 330 is inhibited from being removed from the case 310.

<Fixing pin>
The fixing pin 340 has a knob 341 at its base end, and an elastic portion 342 inserted into the pin holes 315 and 336 of the case 310 and the cover member 330 at its tip, and is formed of a metal wire such as stainless steel. Ru. The elastic portion 342 bulges at the middle portion in the insertion direction, and elastically approaches or separates and deforms.
When the fixing pin 340 is inserted into each pin hole 315, 336, the elastic portion 342 presses the inner surface of the pin hole 315, 336 to prevent the fixing pin 340 from coming off from the pin holes 315, 336.

〔changer〕
The changer 350 for transferring the support 10 accommodated in the cartridge 300 to the cell administration container 20 is provided with a changer main body 351 provided with a plunger pressing projection 356 for pressing the plunger 320 of the cartridge 300 and the changer main body And a lifting member 361 attached to the housing 351.
The changer 350 allows the support 10 to be transferred to the cell delivery container 20 without touching the cell delivery container 20 with the hand of the glove of the isolator.

<Changer body>
The changer main body 351 includes a substantially rectangular base plate 352, a leg member 353 protruding from the lower surface of the base plate 352, an elevation shaft 354 projecting upward from four corners of the base plate 352, and a coil spring through which the elevation shaft 354 is inserted. 355 and a plunger pressing projection 356 that protrudes from an in-plane proper position of the base plate 352 and presses the plunger 320.
The coil spring 355 is disposed below the elevating shaft 354 and is prevented from being separated from the base plate 352. The upper end portion of the elevation shaft 354 is exposed above the upper end edge of the coil spring 355.
Since a space is formed by the leg members 353 below the base plate 352, the movement of the changer main body 351 by the hand wearing the glove becomes easy.

Lifting member
The raising and lowering member 361 is a substantially rectangular flat tray 362, an axial hole 363 formed at four corners of the tray 362 to be penetrated, and a raising and lowering shaft 354 inserted therein, and a plunger 363 formed penetrating in the plane of the tray 362 It has an insertion and withdrawal hole 364 in which the pressing projection 356 is inserted and withdrawn, and a first guide projection 365 and a second guide projection 366 which project upward from the periphery of the insertion and withdrawal hole 364.
The shaft hole 363 has an inner diameter that allows the lift shaft 354 to pass through and the coil spring 355 can not pass through. When the elevating shaft 354 is inserted into the shaft hole 363, the tray 362 is supported in a floating state from the base plate 352 by the elastic force of the coil spring 355, and the plunger pressing projection 356 is positioned below the projecting hole 364. When an external force is applied to the tray 362 to make it approach the base plate 352, the plunger pressing projection 356 protrudes above the tray 362 through the projection hole 364.

  The first guide projection 365 is inserted into both the case 310 of the cartridge 300 and the respective first positioning holes 27 and 312 of the cell administration container 20 in communication with each other. Further, the second guide projection 366 is inserted into both holes in a state where the case 310 of the cartridge 300 and the second positioning holes 28 and 313 of the cell administration container 20 are in communication with each other. On the tray 362 side of the first guide projection 365 and the second guide projection 366, a large diameter portion 365a having a larger diameter than the first positioning holes 27, 312 and the second positioning holes 28, 313, respectively. 366a is formed. The large diameter portions 365a and 366a limit the amount of pressing of the plunger 320 by the plunger pressing projection 356 to a predetermined amount.

<Operating tool>
FIG. 14 is a perspective view showing an example of a manipulation tool for operating the cell administration container.
The operation tool 370 is a tweezer-like jig made of a metal wire rod having a U-shape in plan view.
The tip of the operation rod 371 extending substantially in parallel is formed in a crank shape in the same direction, and is inserted into the operation rod insertion hole 30 of the cell administration container 20 at the foremost end of the operation rod 371. It has an insertion rod 372a, 372b.
The manipulation tool 370 is elastically deformable, and when an external force causing the manipulation rod portion 371 to approach and deform is applied and then the external force is removed, the two manipulation rod portions 371 return to the original posture by elasticity.
Since the distance between the insertion rod portions 372 in the state where no external force is applied is wider than the distance between the operation rod insertion holes 30, each insertion rod portion 372 is inserted in the two operation rod insertion holes 30, each insertion rod portion Since 372 pushes the operating rod insertion holes 30 away from each other by elastic force, the operating tool 370 is effectively prevented from coming off from the operating rod insertion holes 30.
Further, the axial lengths of the two insertion rod portions 372a and 372b are different. A long insertion rod portion 372a is first inserted into one operation rod insertion hole 30, and a short insertion rod portion 372b can be inserted into the other operation rod insertion hole 30 later, I improve the sex.

<Method of transferring support>
A method of transferring the support 10 from the cartridge 300 to the cell administration container 20 will be described based on FIG. 13 and FIG. Fig.15 (a), (b) is a front view which shows operation | movement of a changer.
First, the tray 362 is placed on the changer body 351. The tray 362 is supported by the coil spring 355 in a state of being lifted from the base plate 352 of the changer body 351.
The cartridge 300 with the fixing pin 340 attached is set in the tray 362. The first guide projection 365 is inserted into the first positioning hole 312 and the second guide projection 366 is inserted into the second positioning hole 313. The first and second guide protrusions 365, 366 and the first and second positioning holes 312, 313 allow the cartridge 300 to be installed on the tray 362 with the plunger 320 projecting downward. In this state, by removing the fixing pin 340, the connection between the cover member 330 and the case 310 is released.
By operating the operation piece 335 of the cover member 330, the cover member 330 is pulled out along the longitudinal direction of the case 310. Thereby, the support 10 accommodated in the support accommodation hole 311 is exposed.

The cell administration container 20 is stacked and mounted on the case 310 with the slit 23 side facing upward. At this time, the insertion rod portions 372 of the two operation tools 370 are inserted into the operation rod insertion holes 30 from the respective longitudinal ends of the cell administration container 20 so that the cell administration container 20 is not in direct contact with them. Maintain sterile condition. The first guide projection 365 is inserted into the first positioning hole 27 and the second guide projection 366 is inserted into the second positioning hole 28. By the above-described operation, as shown in FIG. 15A, the support accommodation hole 311 of the case 310 and the support accommodation recess 22 of the cell administration container 20 face each other.
In this state, when the cell administration container 20 is pressed downward by the operation tool 370, the tray 362 descends against the elastic biasing force of the coil spring 355 as shown in FIG. Projecting upward from the tray 362 from 364, the plunger 320 is pressed. As the plunger 320 is pressed by the plunger pressing projection 356, the support 10 in the case 310 is pushed up and moved into the support housing recess 22.

<Removal stand>
A fixture and method for removing only the cell administration container from the cartridge and tray will be described. FIG. 16 (a) is a three-sided view showing a removal stand for removing the cell administration container, and FIG. 16 (b) is a three-sided view showing an operation jig.
The removal base 380 includes a pedestal 381 having a substantially rectangular shape in top view, and a wall 391 rising upward from the pedestal 381 at one end in the longitudinal direction of the pedestal 381.
The pedestal 381 has a rail-shaped first groove 382 extending in the longitudinal direction, and a second groove 383 formed in the first groove 382 in the longitudinal direction. The cell administration container 20 is fitted to the first groove 382, and the projection 25 of the cell administration container 20 is fitted to the second groove 383.

First and second insertion holes 384 and 385 are formed in the first groove portion 382 of the pedestal 381 so as to penetrate in the thickness direction. In the first and second insertion holes 384 and 385, the first guide protrusion 365 and the second guide protrusion 366 of the lifting member 361 having the cell administration container 20 mounted thereon are respectively inserted.
The wall portion 391 is formed with an operation rod insertion hole 392 extending in the longitudinal direction of the removal stand 380. The operating rod insertion hole 392 is formed at a position which is aligned with the operating rod insertion hole 30 of the cell administration container 20 when the cell administration container 20 is fitted to the first groove portion 382.

<< Operation jig >>
The operation jig 400 includes a grip portion 401 which is fitted in the first groove portion 382 and gripped by the operator, and two operation rods 402 a and 402 b which linearly project from the grip portion 401. One operating rod 402a is long, and the other operating rod 402b is shorter than the operating rod 402a. Similar to the insertion rod portions 372a and 372b of the operation tool 370 shown in FIG. 14, the operability of the operation jig 400 is improved by making the lengths of the operation rods 402a and 402b different.

<< removal method >>
After the state shown in FIG. 15 (b) is reached, the lifting member 361 with the case 310 and the cell administration container 20 attached is removed from the changer main body 351. Due to the frictional force between the case 310 and the cell administration container 20 and the first and second guide protrusions 365 and 366 of the elevation member 361, the case 310 and the cell administration container 20 do not fall even if they are turned upside down.
The first and second guide protrusions 365, 366 are respectively inserted into the first and second insertion holes 384, 385 of the removal stand 380. At this time, the cell administration container 20 is fitted in the first groove 382, and the projection 25 of the cell administration container 20 is fitted in the second groove 383.
Insert the operating rods 402 a and 402 b of the operating jig 400 into the operating rod insertion hole 30, and insert the tips of the operating rods 402 a and 402 b exposed from the operating rod insertion hole 30 into the operating rod insertion hole 392. The cell administration container 20 is scab-like.
If the lifting member 361 is lifted in this state, the case 310 is lifted while being held by the lifting member 361 by the frictional force with the first and second guide protrusions 365 and 366, and only the cell administration container 20 is a pedestal Remain on 381.

As described above, by using the removal table, the cell administration container 20 can be removed from the case 310 without touching the cell administration container 20 with the hand equipped with the glove of the isolator.
Thereafter, using the operation jig 400, the cell administration container 20 removed from the elevating member 361 is installed on the container pedestal 40 shown in FIG. 4 and the necessary work is performed.

[Example of embodiment of the present invention, action, summary of effects]
First Embodiment
The cell administration device 100 according to the present embodiment has a support housing recess 22 formed on the upper surface 21 and a slit 23 penetrating the bottom wall of the support housing recess, and is porous in the support housing recess. Cell administration container 20 for containing the stromal cell support 10, a cell suspension feeder 130 for supplying a cell suspension to the porous cell support housed in the support accommodation recess, a porous cell support And a water absorbing material contacting means 190 for bringing the water absorbing material P for absorbing water through the slit into contact with the slit from below.
According to this aspect, since the manufacture of the support on which the cells are seeded can be mechanized, a large amount of support can be manufactured in a short time.

Second Embodiment
In the present embodiment, the volume of the support housing recess 22 is characterized in that it is at least approximately equal to or larger than the sum of the volume of the porous cell support 10 and the volume of the liquid that can be impregnated by the porous cell support. Do.
According to this aspect, the porous cell support impregnated with the induction aqueous solution is accommodated in the support accommodating recess, and the whole cell suspension is absorbed by the support on the accommodated porous cell support. Can meet.

Third Embodiment
In the present embodiment, the porous cell support 10 has a communicating small pore structure having an average pore diameter of 200 to 300 μm, and is connected to the outwardly domed curved surface 11 and the edge connected to the edge of the curved surface The inner surface of the support housing recess 22 is a block-like shape having at least a part of the curved surface when the porous support is accommodated with the flat surface as the upper surface. It is characterized in that it has a curved shape that contacts.
According to this aspect, it is possible to allow the cell suspension to penetrate smoothly into the support by utilizing the power of the water-absorbent material to suck the induction aqueous solution.

Fourth Embodiment
In the present embodiment, the support housing recess 22 is characterized by having a curved inner surface 24 which is convex downward, and the slit 23 is formed at a position corresponding to the top of the curved surface.
According to this aspect, by forming the slit at the lowermost part of the support housing recess, it is possible to absorb the induced aqueous solution smoothly by using the attraction.

Fifth Embodiment
The cell administration apparatus 100 according to the present embodiment includes a plurality of cell administration containers 20, a plurality of water absorbent material contact / separation means 190 provided corresponding to each cell administration container, and a plurality of cell suspension supply devices 130. And a cell suspension feeder driving means 160 driven across the dose container.
According to this aspect, by mechanizing the complicated cell administration operation, the product can be homogenized as compared with the case of manufacturing the support by human operation. In addition, since production of a support on which cells are seeded can be mechanized, a large amount of support can be produced in a short time.

  P: water absorbing material, 10: support, 20: cell administration container, 22: support accommodation recess, 23: slit, 40: container holder, 41: base body, 42: through hole, 48: handle portion, 50 ... Syringe, 60 ... cell mixer, 80 ... stabilizer, 100 ... cell administration device, 110 ... cell administration container installation part, 130 ... cell suspension feeder, 160 ... cell suspension feeder driving means, 161 ... stage 170: Water absorbing material holding member, 180: Cushion material, 190: Water absorbing material contact / separation means, 201: Rotating shaft member, 202: Pressed member, 205: Air cylinder, 300: Cartridge, 320: Plunger, 330: Cover Members, 340: Fixing pins, 350: Changers, 351: Changer bodies, 361: Lifting members, 370: Operation tools, 380: Removal bases, 400: Operation jigs

Claims (5)

A cell administration container having a support accommodating recess formed on the upper surface and a slit penetrating the bottom wall of the support accommodating recess, the porous cell support being accommodated in the support accommodating recess; ,
A cell suspension feeder for supplying a cell suspension to a porous cell support housed in the support housing recess;
A water absorbing material contacting means for bringing a water absorbing material, which absorbs the aqueous solution into which the porous cell support has been impregnated, through the slit from above and below the slit, Administration device. Volume of the support housing recess, wherein at least said porous cell support volume and the porous cell support sum and whether such a volume of liquid which can be impregnated in, greater than this The cell administration device according to claim 1. The porous cell support has an average pore diameter have the continuous small hole structure of 200-300 [mu] m, has a curved surface that bulges in a dome shape outward in a part,
The inner surface of the support housing recess is a curved shape that is in surface contact with at least a part of the curved surface when the porous support is housed with the curved surface as the lower surface. The cell administration device according to 2.   The said support accommodating recess has a curved inner surface which becomes convex downward, The said slit is formed in the position corresponding to the top part of the said inner surface, The any one of the Claims 1 thru | or 3 characterized by the above-mentioned. The cell administration device as described in. A plurality of said cell administration containers,
A plurality of the water absorbent material contacting means provided corresponding to the respective cell administration containers;
5. A cell suspension feeder driving means for driving the cell suspension feeder across a plurality of the cell administration containers, the method according to any one of claims 1 to 4, further comprising: The cell administration device as described in.

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