JP2020162457A - Cell culture device - Google Patents

Abstract

To improve the adhesion property of two members that sandwich a culture membrane in an assembled state.SOLUTION: A cell culture device has a culture membrane having a membrane body and a support frame joined to the membrane body, a first member comprising a first cylindrical part extending in a cylindrical shape in the axial direction, and a loading part which is provided with an axial end face facing the axial direction and on which the culture membrane is loaded in contact with the support frame, and a second member comprising a second cylindrical part extending axially in a cylindrical shape and a facing part that faces the axial end face of the loading part in the axial direction and sandwiches the support frame together with the loading part therebetween. The first member and the second member have a locking part that regulates relative movement in the axial direction in which the loading part and the facing part are assembled from each other so as to sandwich the support frame and are separated from each other.SELECTED DRAWING: Figure 1

Description

The present invention relates to a cell culture device used for culturing cells.

Conventionally, a cell culture device for culturing cells is known (for example, Patent Document 1). The cell culture device described in Patent Document 1 includes a culture membrane, an outer cylinder, and an inner cylinder. The outer cylinder has a slit extending in the axial direction, and the presence of the slit allows elastic deformation in the radial direction. The inner cylinder has a mounting portion on which the culture membrane is placed, and has an outer diameter larger than the inner diameter of the outer cylinder. The inner cylinder is axially moved relative to the outer cylinder in a state where the culture membrane is placed, and is inserted into the outer cylinder from the mounting side. When this insertion is performed, the outer cylinder elastically deforms outward in the radial direction, so that the inner cylinder and the outer cylinder are in contact with each other without a gap in the radial direction, and the culture membrane is sandwiched between the inner cylinder and the outer cylinder. Be retained.

Japanese Unexamined Patent Publication No. 2017-29092

However, in the above cell culture device, the holding force of the culture membrane between the inner cylinder and the outer cylinder depends on the frictional force which is the elastic force between the inner cylinder and the outer cylinder. Therefore, in this cell culture device, if the above-mentioned elastic force is small, the adhesion between the inner cylinder and the outer cylinder is lowered during culturing, and the relative movement of the inner cylinder and the outer cylinder in the axial direction occurs after assembly. , The sealing property between both sides of the culture membrane is reduced.

The present invention has been made in view of such a point, and by improving the adhesion in the assembled state of the two members sandwiching the culture membrane, the sealing property between both sides of the culture membrane is sandwiched. It is an object of the present invention to provide a cell culture device capable of ensuring.

The cell culture apparatus according to one aspect of the present invention includes a culture membrane having a membrane body, a support frame joined to the membrane body, a first tubular portion extending axially in a tubular shape, and an axially oriented cell culture device. A first member having a mounting portion on which the culture film is placed in contact with the support frame on which the axial end face was formed, a second tubular portion extending axially in a tubular shape, and a front portion. A cell culture device comprising a second member that is axially opposed to the axial end face of the description placement portion and has an opposite portion that sandwiches the support frame with the description placement portion. The one member and the second member have a locking portion that regulates relative movement in the axial direction in which the above-mentioned placing portion and the facing portion are assembled from each other so as to sandwich the support frame and are separated from each other. , A cell culture device.

According to this configuration, the first member and the second member are assembled by the locking portion in a state where the mounting portion of the first member and the facing portion of the second member are assembled to each other so as to sandwich the support frame of the culture membrane. It is possible to regulate the relative movement in the axial direction away from each other with the member. Therefore, it is possible to improve the adhesion between the first member and the second member that sandwich the culture membrane in the assembled state, and it is possible to secure the sealing property between both sides of the culture membrane during cell culture. it can.

It is an exploded perspective view of the cell culture apparatus which concerns on 1st Embodiment. It is a side view (note that a part is a cross-sectional view) of the cell culture apparatus of 1st Embodiment. It is a top view of the cell culture apparatus of the first embodiment. FIG. 5 is a cross-sectional view of the cell culture device of the first embodiment cut at IV-IV shown in FIG. It is a figure for demonstrating the locking part of the 1st member and the 2nd member provided in the cell culture apparatus of 1st Embodiment. It is a figure for demonstrating the locking part of the 1st member and the 2nd member provided in the cell culture apparatus which concerns on one modified form. It is a side view (left half) and sectional view (right half) of the cell culture apparatus which concerns on 2nd Embodiment. It is a figure showing the state before locking of the locking part of the 1st member and the 2nd member provided in the cell culture apparatus which concerns on 3rd Embodiment. It is a figure showing the state after locking of the locking part of the 1st member and the 2nd member provided in the cell culture apparatus which concerns on 3rd Embodiment.

Hereinafter, specific embodiments of the cell culture device according to the present invention will be described with reference to the drawings.

[First Embodiment]
The cell culture device 1 of the first embodiment is a device for culturing cells having a scaffold dependence. The cell culture device 1 can culture two types of cells on both sides of the membrane. As shown in FIGS. 1, 2, 3, and 4, the cell culture instrument 1 includes a culture membrane 10, a first member 20, and a second member 30. The cell culture instrument 1 sandwiches and fixes the culture membrane 10 between the first member 20 and the second member 30.

The culture membrane 10 is a sheet-like membrane in which cells are cultured. The culture membrane 10 has a membrane body 11 and a support frame 12. The membrane body 11 is a membrane body portion that serves as a scaffold for cells during cell culture. The membrane body 11 is a porous membrane having pores on both sides. The film body 11 is formed of, for example, a thermosetting resin such as polyurethane, silicone resin, phenol resin, epoxy resin, unsaturated polyester, or polyimide. The film thickness of the film body 11 is, for example, 0.1 μm-100 μm.

The support frame 12 is a base material that supports and reinforces the film body 11. The support frame 12 is an annular frame having a circular hole 12a in the center. The support frame 12 is joined and fixed to the peripheral edge of the film body 11. The shape of the support frame 12 may include at least an annular frame portion, and may further include a protrusion protruding radially outward from a part of the frame portion in the circumferential direction. Good. This protrusion has a function for facilitating handling when, for example, a person grips the culture membrane 10 with a jig such as tweezers. The support frame 12 is made of a material harder than the film body 11. The material of the support frame 12 is, for example, a thermoplastic resin such as polyethylene, polypropylene, ABS resin, acrylic resin, polyethylene terephthalate (PET), or a thermosetting elastomer such as silicone rubber. The thickness of the support frame 12 is, for example, 0.05 mm to 0.5 mm.

The first member 20 is a base material for sandwiching and fixing the culture film 10. The first member 20 is a member formed in a tubular shape. The first member 20 is made of a resin material, rubber, or the like. The first member 20 has a first tubular portion 21, a mounting portion 22, and a side opening portion 23. The first cylinder portion 21 and the mounting portion 22 are integrally formed with each other.

The first tubular portion 21 is a hollow portion extending in a tubular shape in the axial direction X. The first tubular portion 21 is formed, for example, in a cylindrical shape. The inner diameter of the first tubular portion 21 may be constant regardless of the position in the axial direction X, but changes according to the position in the axial direction X (specifically, the shaft far from the mounting portion 22). It may be larger as the position in the direction X). The first tubular portion 21 has a peripheral wall portion 21a. The inner peripheral surface of the peripheral wall portion 21a is formed so that the culture film 10 can be guided to the mounting portion 22 in the axial direction X.

The peripheral wall portion 21a has a notch portion 21b. The notch portion 21b is provided so as to penetrate the peripheral wall portion 21a in the radial direction and extend in the axial direction X. The cutout portion 21b extends in the axial direction X from the axial end portion where the culture film 10 in the first tubular portion 21 is inserted into the first member 20 in the axial direction X to the portion where the mounting portion 22 is provided. .. The cutout portion 21b is provided in a part of the peripheral wall portion 21a in the circumferential direction, and is provided in one place in the peripheral wall portion 21a. The cutout portion 21b is a slit for inserting a jig for attaching / detaching the culture membrane 10 sandwiched between the first member 20 and the second member 30 to / from the mounting portion 22 from the outside in the radial direction. That is, the notch portion 21b has a function of enabling the jig inserted from the outside in the radial direction with respect to the first member 20 to be movable in the axial direction X.

The mounting portion 22 is a portion on which the culture membrane 10 is placed. The mounting portion 22 is connected to one end of the first tubular portion 21 in the axial direction. The mounting portion 22 is formed in an annular shape on the inner peripheral side of one end in the axial direction of the first tubular portion 21. The mounting portion 22 may be provided intermittently in the circumferential direction on the inner peripheral side of one end of the first tubular portion 21 in the axial direction. The mounting portion 22 has a wall portion 22a on which an axial end surface facing the axial direction X is formed, and a hole portion 22b formed in a circular shape in the central portion of the wall portion 22a. The mounting portion 22 abuts on the support frame 12 of the culture film 10 at the inner axial end surface of the wall portion 22a to hold the culture film 10. The culture film 10 held by the wall portion 22a of the mounting portion 22 is exposed to the outside through the hole portion 22b of the mounting portion 22.

The side surface opening 23 is provided so as to penetrate the peripheral wall portion 21a of the first tubular portion 21 in the radial direction. The side opening 23 is a hole for communicating the inside of the first cylinder 21 to the outside. The side surface opening 23 is provided in a part of the peripheral wall portion 21a in the circumferential direction, and is provided at a predetermined portion of the peripheral wall portion 21a. The predetermined portion where the side opening portion 23 is provided includes at least a portion on the mounting portion 22 side in the axial direction X with respect to the culture membrane 10 mounted on the mounting portion 22. The side opening 23 has an air bleeding function for discharging the air accumulated on the lower surface of the culture film 10 placed on the mounting portion 22 to the outside. The side opening 23 is provided at the same position in the first member 20 as the notch 21b in the circumferential direction. The side opening 23 is continuously arranged in the axial direction X with respect to the notch 21b, and is integrated with the notch 21b.

The second member 30 is a base material for sandwiching and fixing the culture film 10 which is paired with the first member 20. The second member 30 is a member formed in a tubular shape. The second member 30 is inserted and arranged in the first tubular portion 21 of the first member 20. The second member 30 is made of a resin material, rubber, or the like. The second member 30 has a second tubular portion 31, an opposing portion 32, and a grip portion 33. The second cylinder portion 31, the facing portion 32, and the grip portion 33 are integrally formed with each other.

The second tubular portion 31 is a hollow portion extending in a tubular shape in the axial direction X. The second tubular portion 31 is formed, for example, in a cylindrical shape. The outer diameter of the second tubular portion 31 has a size substantially matching the inner diameter of the first tubular portion 21 of the first member 20. The inner diameter or outer diameter of the second tubular portion 31 may be constant regardless of the position in the axial direction X, but changes according to the position in the axial direction X (specifically, the mounting portion 22). The position in the axial direction X farther from is larger).

The facing portion 32 is a portion facing the wall portion 22a of the mounting portion 22 of the first member 20 in the axial direction X. The facing portion 32 has a function of sandwiching the culture film 10 with the mounting portion 22 of the first member 20. The facing portion 32 is connected to one end in the axial direction of the second tubular portion 31. The facing portion 32 is formed in an annular shape on the inner peripheral side of one end in the axial direction of the second tubular portion 31. The facing portion 32 may be provided intermittently in the circumferential direction on the inner peripheral side of one end in the axial direction of the second tubular portion 31.

The facing portion 32 has a wall portion 32a on which an axial end surface facing the axial direction X is formed, and a hole portion 32b formed in a circular shape in the central portion of the wall portion 32a. The facing portion 32 abuts on the support frame 12 of the culture film 10 at the outer axial end surface of the wall portion 32a and holds the culture film 10. The culture film 10 sandwiched between the mounting portion 22 and the facing portion 32 is exposed to the inner space of the second tubular portion 31 through the hole portion 32b of the facing portion 32.

The grip portion 33 has a function for facilitating the handling of the second member 30 when a person assembles the second member 30 to the first member 20 with a jig such as tweezers. The grip portion 33 has an arm portion 33a and a connecting portion 33b. A plurality (for example, three) arm portions 33a are provided. One end of each arm portion 33a is connected to the other end in the axial direction opposite to the facing portion 32 in the second tubular portion 31. The arm portions 33a are arranged at intervals in the circumferential direction of the second tubular portion 31. The arm portion 33a extends outward in the axial direction X from the other end in the axial direction of the second tubular portion 31. The connecting portion 33b is formed in an annular shape so as to connect the other ends of the plurality of arm portions 33a. The connecting portion 33b is notched in a part in the circumferential direction and is formed in a C shape.

The first member 20 and the second member 30 have a locking portion that regulates relative movement in the axial direction X that is separated from each other in the assembled state. That is, the first member 20 has a concave portion 25 as a locking portion, and the second member 30 has a convex portion 35 as a locking portion. As shown in FIG. 5, the concave portion 25 and the convex portion 35 regulate relative movement in the axial direction X, which is separated from each other in the assembled state of the first member 20 and the second member 30.

The recess 25 of the first member 20 is a groove provided in the inner surface of the peripheral wall portion 21a of the first tubular portion 21 so as to be recessed outward in the radial direction. The recesses 25 are provided at two locations (for example, 180 ° different positions) in the circumferential direction of the first member 20, for example. The recess 25 is formed in an L shape. That is, the recess 25 has an axial parallel portion 25a extending in the axial direction X and an orthogonal portion 25b extending in a direction orthogonal to the axial direction X. The axially parallel portion 25a and the orthogonal portion 25b each have a width equal to or larger than the width of the convex portion 35 of the second member 30. The axially parallel portion 25a extends from the other end in the axial direction opposite to the mounting portion 22 in the first tubular portion 21 toward the one end side in the axial direction. The orthogonal portion 25b is connected to one end in the axial direction of the axially parallel portion 25a and extends in the orthogonal direction. The orthogonal portion 25b forms a locking wall 26 on which the convex portion 35 of the second member 30 is locked on the other end side in the axial direction.

The convex portion 35 of the second member 30 is a protruding portion provided on the outer surface of the second tubular portion 31 so as to project outward in the radial direction. The convex portions 35 are provided at two locations (for example, 180 ° different positions) in the circumferential direction of the second member 30, for example. The convex portion 35 is formed in a size corresponding to the concave portion 25 (particularly, the orthogonal portion 25b). Specifically, the convex portion 35 has a width (that is, an axial length) corresponding to the width (that is, the axial length) of the orthogonal portion 25b of the concave portion 25.

The width of the orthogonal portion 25b of the recess 25 may change depending on the position in the circumferential direction. For example, as shown in FIG. 6, the width thereof is such that the convex portion 35 easily moves from the side near the axial parallel portion 25a in the orthogonal portion 25b to the far back side when the first member 20 and the second member 30 are assembled. It may be widest on the side closer to the axis parallel portion 25a and gradually narrower from the axis parallel portion 25a to the far back side. Further, the width is such that the predetermined portion has the minimum width so that it is difficult to move from the predetermined portion to the side closer to the axial parallel portion 25a after the convex portion 35 fits into the predetermined portion on the back side of the orthogonal portion 25b. It may be located on the inner side farther from the axis parallel portion 25a and the width of the predetermined portion may be wider than the minimum width.

The assembly of the first member 20 and the second member 30 is performed so that the convex portion 35 moves along the concave portion 25. The convex portion 35 can be fitted near the tip end side of the concave portion 25 and locked to the locking wall 26. The first member 20 and the second member 30 are in a state of being assembled to each other by fitting the convex portion 35 into the concave portion 25 and being locked to the locking wall 26, and are relative to each other in the axial direction X. Movement will be restricted.

In the state where the first member 20 and the second member 30 are assembled by fitting the convex portion 35 into the concave portion 25, the distance between the mounting portion 22 and the facing portion 32 in the axial direction X is set to culture. The thickness of the support frame 12 of the film 10 is set to be smaller than the thickness before assembly. As a result, in the assembled state, the culture film 10 is sandwiched between the mounting portion 22 of the first member 20 and the facing portion 32 of the second member 30.

Next, the assembly of the cell culture device 1 will be described.

The culture membrane 10, the first member 20, and the second member 30, which are the components of the cell culture instrument 1, are prepared. The cell culture device 1 is assembled manually by a person using a jig such as tweezers. As the assembly procedure, first, the culture membrane 10 is assembled to the first member 20. In assembling the culture membrane 10 to the first member 20, after gripping the culture membrane 10 with a jig, the culture membrane 10 is attached to the first member 20 with respect to the mounting portion 22 on the first member 20. This is realized by moving the culture film 10 in the axial direction X from the opposite side and inserting the culture film 10 so that the culture film 10 is placed in contact with the axial end face of the mounting portion 22 of the first member 20. The culture membrane 10 may be assembled to the first member 20 described above in a state where the first member 20 is set so that the axial end portion on the mounting portion 22 side is located downward. ..

Next, the second member 30 is assembled to the first member 20 on which the culture film 10 is placed on the mounting portion 22 as described above. In this assembly, first, the grip portion 33 of the second member 30 is gripped with a jig, and then the second member 30 is axially oriented with respect to the first member 20 and the mounting portion 22 of the first member 20. The convex portion 35 of the second member 30 is moved along the axially parallel portion 25a of the concave portion 25 of the first member 20 by moving and inserting from the opposite side in the axial direction X toward each other, and then the second member 30 Is moved in one direction in the circumferential direction with respect to the first member 20, and the convex portion 35 of the second member 30 is moved along the orthogonal portion 25b of the concave portion 25 of the first member 20. In the above-mentioned assembly of the second member 30 to the first member 20, the second member 30 is set in a state where the first member 20 is set so that the axial end portion on the mounting portion 22 side is located downward. It may be inserted from above the first member 20.

When the above movement is performed and the convex portion 35 of the second member 30 fits near the tip end side of the concave portion 25 of the first member 20, the axial directions X of both members 20 and 30 move relative to each other. Is regulated and both members 20 and 30 are attached to each other, and the assembly of both members 20 and 30 is completed. When the assembly of both members 20 and 30 is completed, the mounting portion 22 and the facing portion 32 sandwich the culture film 10 (specifically, the support frame 12) between them, and the convex portion 35 of the second member 30 is formed. It is locked to the locking wall 26 of the recess 25 of the first member 20.

According to the structure of the cell culture device 1, after assembling the first member 20 and the second member 30, the support frame 12 of the culture membrane 10 is sandwiched between the first member 20 and the second member 30. The relative movement of the first member 20 and the second member 30 in the axial direction X can be regulated by the concave portion 25 and the convex portion 35 as the locking portions. Therefore, the adhesion between the first member 20 and the second member 30 holding the culture membrane 10 in the assembled state can be improved, and the sealing property between both sides sandwiching the culture membrane 10 during cell culture can be improved. Can be secured. The culture film 10 located in the side opening 23 is not sandwiched between the first member 20 and the second member 30, but since the region is narrow, only the elastic pressing force of the support frame 12 and the second member 30 is used. It is sealed.

After the assembly of the first member 20 and the second member 30 is completed as described above, cell culture is performed on one surface of the culture membrane 10. For cell culture, the cell culture device 1 may be placed in a well of a cell culture plate and filled with a culture solution. In the cell culture on one surface of the culture film 10, first, the cell culture device 1 is immersed in the culture solution so that one surface of the culture film 10 on the second member 30 side faces vertically upward, and the cell culture device 1 is immersed therein. This is done by supplying the first cell to the space on one side of the surface in the state.

As described above, when the culture of the first type of cells is completed, the assembly of the first member 20 and the second member 30 of the cell culture device 1 is then released, the culture membrane 10 is turned upside down, and the first member is again. The first member 20 and the second member 30 are assembled. The disassembly of the first member 20 and the second member 30 is performed in the reverse procedure of the above-mentioned assembly procedure. Specifically, in the disassembly, the second member 30 is first moved to the other side in the circumferential direction with respect to the first member 20, and then the second member 30 is moved to each other in the axial direction X with respect to the first member 20. It is realized by moving in the direction of separation. When this assembly is released, the culture membrane 10 is then moved in the axial direction X by a jig from the state of being mounted on the mounting portion 22 of the first member 20, then turned upside down and again. , It is placed in contact with the axial end face of the mounting portion 22. Then, as described above, the second member 30 is assembled to the first member 20 on which the culture film 10 is placed on the mounting portion 22.

When the above assembly is completed, the mounting portion 22 and the facing portion 32 sandwich the culture membrane 10 between them, and the convex portion 35 of the second member 30 engages with the locking wall 26 of the concave portion 25 of the first member 20. Since it is stopped, an effect similar to the above effect can be obtained. When this assembly is completed, cell culture is performed on the other surface of the culture membrane 10. In the cell culture on the other surface of the culture membrane 10, first, the cell culture instrument 1 is immersed in the culture solution with the other surface of the culture membrane 10 on the second member 30 side facing vertically upward, and the cell culture instrument 1 is immersed therein. This is done by supplying a second type of cell to the space on the other side in the state.

As described above, according to the structure of the cell culture device 1, two types of cells can be cultured on both sides of the culture membrane 10. Further, in culturing two types of cells on both sides of the culture membrane 10, the culture membrane 10 itself is moved up and down after culturing the cells on one side of the culture membrane 10 and after disassembling the first member 20 and the second member 30. It may be inverted, and it is not necessary to invert the first member 20 and the second member 30 themselves. Therefore, workability in culturing two types of cells on both sides of the culture membrane 10 can be improved.

Further, when culturing two types of cells on both sides of the culture membrane 10, it is not necessary to make the cell culture device vertically symmetrical in consideration of the upside down of the culture membrane 10. Therefore, the structure of the mounting portion 22 in the axial direction X can be simplified from the culture membrane 10 mounted on the mounting portion 22 of the first member 20, and the cell culture device 1 can be miniaturized. In order to realize cell culture, it is possible to reduce the size of the well and the amount of the culture solution. Further, since the culture membrane 10 can be independently removed from the first member 20 and the second member 30, the culture membrane 10 after cell culture can be independently evaluated by microscopic observation or the like. Compared with the structure in which the holding member and the holding member are integrated, the evaluation can be easily and appropriately performed.

Further, in the cell culture device 1, the first member 20 has a notch 21b that penetrates the peripheral wall portion 21a in the radial direction and extends in the axial direction X at the first tubular portion 21. The notch 21b is a slit for inserting a jig such as tweezers from the outside in the radial direction of the first member 20 and moving it in the axial direction X. The culture membrane 10 is placed on the mounting portion 22 of the first member 20 and sandwiched between the first member 20 and the second member 30, but the cutout portion is used for attaching / detaching to / from the mounting portion 22. It can be gripped by a jig such as tweezers inserted from 21b. Therefore, the culture film 10 can be easily attached to and detached from the mounting portion 22 of the first member 20 using a jig, and the cell culture device is used to realize cell culture on both sides of the culture film 10. It is not necessary to prepare many parts as No. 1, and cell culture can be performed on both sides of the culture membrane 10 with the minimum configuration.

The first member 20 also has a side opening 23 that penetrates the peripheral wall portion 21a in the radial direction at the first tubular portion 21. The side surface opening 23 is provided at a predetermined portion of the peripheral wall portion 21a including at least a portion on the mounting portion 22 side in the axial direction X with respect to the culture film 10 mounted on the mounting portion 22. Therefore, after culturing the first type of cells on one surface of the culture membrane 10, when the culture membrane 10 is turned upside down and the second type of cells are cultured on the other surface, the first type of cells are cultured. Since the air accumulated on the surface to be cultivated can be discharged to the outside of the first member 20 through the side opening 23, it is possible to prevent the air from remaining on the culture surface of the first seeded cell. It is possible to avoid the inconvenience caused by the residual air (for example, the first type of cells do not come into contact with the culture medium).

Further, the cutout portion 21b and the side surface opening portion 23 are provided at the same positions in the first member 20 in the circumferential direction, and are continuously arranged and integrated in the axial direction X. Therefore, the mold structure of the first member 20 provided with the notch 21b and the side opening 23 can be simplified, and the first member 20 can be manufactured accurately and easily.

In the first embodiment described above, the concave portion 25 of the first member 20 and the convex portion 35 of the second member 30 correspond to the "locking portion" described in the claims.

By the way, in the first embodiment described above, the notch 21b and the side opening 23 of the first member 20 are provided at the same positions in the circumferential direction and are continuously arranged in the axial direction X. However, the present invention is not limited to this, and the notch 21b and the side opening 23 of the first member 20 are provided at different positions in the circumferential direction and discontinuously arranged in the axial direction X. You may. According to the structure of this modified form, it is possible to secure the torsional rigidity of the first member 20 in the circumferential direction as compared with the structure of the first embodiment described above.

Further, in the first embodiment described above, the first member 20 has a recess 25 provided on the inner surface of the peripheral wall portion 21a of the first tubular portion 21 so as to be recessed outward in the radial direction, and the second member 30. However, there is a convex portion 35 provided on the outer surface of the second tubular portion 31 so as to project outward in the radial direction. However, the present invention is not limited to this, and the first member 20 has a convex portion provided on the inner surface of the first tubular portion 21 so as to project inward in the radial direction, and the second member. 30 may have a recess provided on the outer surface of the second tubular portion 31 so as to be recessed inward in the radial direction.

Further, in the first embodiment described above, two types of cells are seeded on both sides of the culture membrane 10. However, the present invention is not limited to this, and three or more types of cells may be seeded on both sides of the culture membrane 10. For example, three types of cells are prepared, two types of cells are seeded on one surface of the culture membrane 10, then the culture membrane 10 is turned over and one type of cells is seeded on the other surface of the culture membrane 10. You may do it.

[Second Embodiment]
The cell culture device 100 of the second embodiment is an device for culturing cells having a scaffold dependence, similarly to the cell culture device 1 of the first embodiment. Hereinafter, in the cell culture device 100, the same components as those of the cell culture device 1 are designated by the same reference numerals, and the description thereof will be omitted or simplified.

As shown in FIG. 7, the cell culture device 100 includes a culture membrane 10, a first member 110, and a second member 120. The cell culture device 100 sandwiches and fixes the culture membrane 10 between the first member 110 and the second member 120.

Each of the first member 110 and the second member 120 is a base material for sandwiching and fixing the culture film 10. The first member 110 and the second member 120 are members formed in a tubular shape, respectively. The first member 110 and the second member 120 are each made of a resin material, rubber, or the like.

The first member 110 has a first tubular portion 111 extending in a tubular shape in the axial direction X, a mounting portion (not shown), and a side opening 112 that penetrates the first tubular portion 111 in the radial direction. are doing. The side opening 112 is provided at a predetermined portion of the first tubular portion 111 including at least a portion on the mounting portion side (lower side in FIG. 7) in the axial direction X with respect to the culture membrane 10 mounted on the mounting portion. ing. The mounting portion may be an axial end surface of the first tubular portion 111 facing the axial direction X.

The second member 120 is extrapolated and arranged in a part of the first tubular portion 111 of the first member 110. The second member 120 has a second tubular portion 121 extending in a tubular shape in the axial direction X, an opposing portion 122 facing the mounting portion of the first member 110 in the axial direction X, and the first tubular portion 111 in the radial direction. It has a side opening 123 that penetrates. The facing portion 122 is formed in an annular shape on the inner peripheral side of one end (lower end in FIG. 7) in the axial direction of the second tubular portion 121. The facing portion 122 abuts on the support frame 12 of the culture film 10 at the axial end face and holds the culture film 10. The facing portion 122 may be an axial end face of the second tubular portion 121 facing the axial direction X.

The side opening 123 is provided in a part of the second cylinder portion 121 in the circumferential direction, and is provided at a predetermined portion of the second cylinder portion 121. The side opening 123 communicates with the side opening 112 of the first member 110. The predetermined portion where the side opening 123 is provided includes at least a portion on the mounting portion side (lower side in FIG. 7) in the axial direction X with respect to the culture membrane 10 mounted on the mounting portion. The side opening 123, together with the side opening 112 of the first member 110, has an air bleeding function for discharging air accumulated on the lower surface of the culture film 10 placed on the mounting portion to the outside.

The first member 110 and the second member 120 have a locking portion that regulates relative movement in the axial direction X that is separated from each other in the assembled state. That is, the first member 110 has a convex portion 113 as a locking portion, and the second member 120 has a concave portion 124 as a locking portion. The convex portion 113 and the concave portion 124 regulate the relative movement in the axial direction X separated from each other in the assembled state of the first member 110 and the second member 120.

The convex portion 113 is a protruding portion provided on the outer surface of the first tubular portion 111 so as to project outward in the radial direction. The convex portions 113 are provided at two locations (for example, 180 ° different positions) in the circumferential direction of the first member 110, for example. The convex portion 113 is formed in a size corresponding to the concave portion 124. Specifically, the convex portion 113 has a width corresponding to the width of the concave portion 124.

The recess 124 is a groove provided in the inner surface of the second tubular portion 121 so as to be recessed outward in the radial direction. The recess 124 is formed at one end in the axial direction of the second tubular portion 121. The recesses 124 are provided at two locations (for example, 180 ° different positions) in the circumferential direction of the second member 120, for example. The recess 124 is formed so as to be inclined with respect to the axial direction X. That is, the recess 124 is spirally formed so as to extend in the axial direction X while extending in the circumferential direction. The innermost portion of the concave portion 124 forms a locking wall in which the convex portion 113 of the first member 110 is locked.

The assembly of the first member 110 and the second member 120 is performed so that the convex portion 113 moves along the concave portion 124. The convex portion 113 can be fitted into the innermost portion of the concave portion 124 and locked to the locking wall. The first member 110 and the second member 120 are in a state of being assembled to each other by fitting the convex portion 113 into the concave portion 124 and being locked to the locking wall, and move relative to each other in the axial direction X. Will be regulated.

Next, the assembly of the cell culture device 100 will be described.

The culture membrane 10, the first member 110, and the second member 120, which are the components of the cell culture device 100, are prepared. The cell culture device 100 is assembled manually by a person using a jig such as tweezers. As the assembly procedure, first, the culture membrane 10 is assembled to the first member 110. To assemble the culture membrane 10 to the first member 110, after grasping the culture membrane 10 with a jig, the culture membrane 10 is moved in the axial direction X with respect to the first member 110, and the culture membrane 10 is assembled. It is realized by mounting the first member 110 in contact with the axial end surface of the mounting portion. The culture film 10 may be assembled to the first member 110 in a state where the first member 110 is set so that the axial end portion on the mounting portion side is located upward.

Next, the second member 120 is assembled to the first member 110 on which the culture membrane 10 is placed as described above. In this assembly, first, the second member 120 is moved with respect to the first member 110 in the axial direction X closer to each other to be extrapolated and rotated in the circumferential direction, and the convex portion of the first member 110 is assembled. This is achieved by moving 113 along the recess 124 of the second member 120. In the above-mentioned assembly of the second member 120 to the first member 110, the second member 120 is set with the first member 110 set so that the axial end on the mounting portion side is located upward. It may be inserted from above one member 110.

When the above movement is performed and the convex portion 113 of the first member 110 fits into the innermost portion of the concave portion 124 of the second member 120, the axial directions X of both members 110 and 120 move relative to each other in a direction away from each other. Is regulated and both members 110 and 120 are mounted on each other, and the assembly of both members 110 and 120 is completed. When the assembly of both members 110 and 120 is completed, the mounting portion and the facing portion 122 sandwich the culture membrane 10 (specifically, the support frame 12) between them, and the convex portion 113 of the first member 110 becomes the first. It is locked to the locking wall of the recess 124 of the two members 120.

According to the structure of the cell culture device 100, after assembling the first member 110 and the second member 120, the support frame 12 of the culture membrane 10 is sandwiched between the first member 110 and the second member 120, and the first member 110 and the second member 120 are sandwiched between the first member 110 and the second member 120. The relative movement of the one member 110 and the second member 120 in the axial direction X can be regulated by the convex portion 113 and the concave portion 124 as the locking portions. Therefore, the adhesion between the first member 110 and the second member 120 holding the culture membrane 10 in the assembled state can be improved, and the sealing property between both sides sandwiching the culture membrane 10 during cell culture can be improved. Can be secured.

After the assembly of the first member 110 and the second member 120 is completed as described above, cell culture is performed on both sides of the culture membrane 10 by the same method as in the case of the first embodiment. Therefore, according to the cell culture device 100, the same effect as that of the cell culture device 1 of the first embodiment can be obtained.

In the second embodiment described above, the convex portion 113 of the first member 110 and the concave portion 124 of the second member 120 each correspond to the "locking portion" described in the claims.

By the way, in the above-mentioned second embodiment, the first member 110 has a convex portion 113 provided on the outer surface of the first tubular portion 111 so as to project outward in the radial direction, and the second member 120 has a convex portion 113. It has a recess 124 provided on the inner surface of the second tubular portion 121 so as to be recessed outward in the radial direction. However, the present invention is not limited to this, and the first member 110 has a recess provided on the outer surface of the first tubular portion 111 so as to be recessed inward in the radial direction, and the second member 120 is provided. , The inner surface of the second tubular portion 121 may have a convex portion provided so as to project inward in the radial direction.

[Third Embodiment]
The cell culture device 200 of the third embodiment is an device for culturing cells having a scaffold dependence, similar to the cell culture device 1 of the first embodiment. Hereinafter, in the cell culture device 200, the same components as those of the cell culture device 1 are designated by the same reference numerals, and the description thereof will be omitted or simplified.

As shown in FIGS. 8 and 9, the cell culture apparatus 200 includes a culture membrane 10, a first member 210, and a second member 220. The cell culture device 200 sandwiches and fixes the culture membrane 10 between the first member 210 and the second member 220.

Each of the first member 210 and the second member 220 is a base material for sandwiching and fixing the culture film 10. The first member 210 and the second member 220 are members formed in a tubular shape, respectively. The first member 210 and the second member 220 are each made of a resin material, rubber, or the like.

The first member 210 includes a first tubular portion 211 extending in a tubular shape in the axial direction X and a culture membrane 10 similar to the first tubular portion 21 and the mounting portion 22 of the first member 20 of the first embodiment. It has a mounting portion 212 to be mounted. The second member 220 is inserted and arranged in the first cylinder portion 211 of the first member 210, similar to the second cylinder portion 31 and the facing portion 32 of the second member 30 of the first embodiment. The second member 220 has a second tubular portion 221 extending in a tubular shape in the axial direction X, and an opposing portion 222 facing the mounting portion 212 of the first member 210 in the axial direction X.

The first member 210 and the second member 220 have a locking portion that regulates relative movement in the axial direction X that is separated from each other in the assembled state. This locking portion is a so-called snap-fit type locking portion. That is, the first member 210 has an elastically deformed portion 213 as a locking portion, and the second member 220 has a groove portion 223 as a locking portion. The elastically deformed portion 213 and the groove portion 223 regulate the relative movement of the first member 210 and the second member 220 in the axial direction X which are separated from each other in the assembled state.

The elastically deformed portion 213 is a portion extending in a plate shape from the axial end surface of the first tubular portion 211 in the axial direction X. The elastically deformed portion 213 can be elastically deformed in the radial direction. The elastically deformed portions 213 are provided at two locations (for example, 180 ° different positions) in the circumferential direction of the first member 210, for example. A claw portion 214 is provided at the axial tip portion of the elastically deformed portion 213. The claw portion 214 is a hook portion that protrudes inward in the radial direction from the plate-shaped main body of the elastically deformed portion 213. The radial inner surface of the claw portion 214 forms an inclined surface 215 inclined in a tapered shape. The radial width of the claw portion 214 is set to be maximum on the root side and gradually decrease toward the axial tip.

The groove portion 223 is provided on the outer surface of the second tubular portion 221 so as to be recessed inward in the radial direction. The groove portion 223 is formed in the middle of the second tubular portion 221 in the axial direction. The groove portion 223 is a portion on which the claw portion 214 is caught when the first member 210 and the second member 220 approach a predetermined positional relationship in the axial direction X. Grooves 223 are provided at two locations (for example, 180 ° different positions) in the circumferential direction of the second member 220, for example. The groove portion 223 is formed in a size corresponding to the claw portion 214 of the elastically deformed portion 213 of the first member 210. The groove portion 223 forms a locking wall in which the claw portion 214 of the first member 210 is caught and locked.

Assembling of the first member 210 and the second member 220 is performed by relatively moving the claw portion 214 of the elastically deformed portion 213 in the axial direction X so as to be hooked on the groove portion 223, as shown in FIGS. 8 and 9. The first member 210 and the second member 220 are in a state of being assembled to each other when the claw portion 214 is hooked on the groove portion 223 and locked, and the relative movement in the axial direction X separated from each other is restricted. It will be.

Next, the assembly of the cell culture device 200 will be described.

The culture membrane 10, the first member 210, and the second member 220, which are the components of the cell culture instrument 200, are prepared. The cell culture device 200 is assembled manually by a person using a jig such as tweezers. As the assembly procedure, first, the culture membrane 10 is assembled to the first member 210. To assemble the culture membrane 10 to the first member 210, the culture membrane 10 is gripped with a jig, the culture membrane 10 is moved in the axial direction X with respect to the first member 210, and the culture membrane 10 is assembled. This is achieved by mounting the first member 210 in contact with the axial end face of the mounting portion 212. The culture membrane 10 may be assembled to the first member 210 in a state where the first member 210 is set so that the axial end portion on the mounting portion side is located upward.

Next, the second member 220 is assembled to the first member 210 on which the culture membrane 10 is placed on the mounting portion as described above. This assembly is realized by moving the second member 120 with respect to the first member 110 in the axial direction X close to each other and inserting the second member 120 with a jig. Specifically, first, the second tubular portion 221 of the second member 220 is brought into contact with the inclined surface 215 of the claw portion 214 of the elastically deformed portion 213 of the first member 210, and the axial tip of the elastically deformed portion 213 is brought into contact with the inclined surface 215. The portion is flexed outward in the radial direction to be deformed, and then, when the claw portion 214 of the elastic deformed portion 213 reaches the groove portion 223 of the second member 220, the axial tip portion of the elastic deformed portion 213 is returned inward in the radial direction. It is deformed and the claw portion 214 is hooked on the groove portion 223. In the above-mentioned assembly of the second member 220 to the first member 210, the second member 220 is set in a state where the first member 210 is set so that the axial end portion on the mounting portion 212 side is located downward. It may be inserted from above the first member 210.

When the above movement is performed and the claw portion 214 of the elastically deformed portion 213 of the first member 210 is caught in the groove portion 223 of the second member 220, the axial directions X of both members 210 and 220 are relative to each other. The movement is restricted and both members 210 and 220 are attached to each other, and the assembly of both members 210 and 220 is completed. When the assembly of both members 210 and 220 is completed, the mounting portion 212 and the facing portion 222 sandwich the culture film 10 (specifically, the support frame 12) between them, and the claw portion 214 of the first member 210 is inserted. It is locked to the groove 223 of the second member 220.

According to the structure of the cell culture device 200, after assembling the first member 210 and the second member 220, the support frame 12 of the culture membrane 10 is sandwiched between the first member 210 and the second member 220, and the first member 210 and the second member 220 are sandwiched between the first member 210 and the second member 220. The relative movement of the one member 210 and the second member 220 in the axial direction X can be regulated by the elastically deformed portion 213 and the groove portion 223 as locking portions. Therefore, it is possible to improve the adhesion between the first member 210 and the second member 220 that sandwich the culture membrane 10 in the assembled state, and to provide a sealing property between both sides of the culture membrane 10 during cell culture. Can be secured.

After the assembly of the first member 210 and the second member 220 is completed as described above, cell culture is performed on both sides of the culture membrane 10 by the same method as in the case of the first embodiment. Therefore, according to the cell culture device 200, the same effect as that of the cell culture device 1 of the first embodiment can be obtained. Further, according to the cell culture device 200, the assembly of the first member 210 and the second member 220 can be realized only by relatively moving both members 210 and 220 in the axial direction X, which simplifies the assembling property. Therefore, the assembly work can be facilitated.

In order to release the assembly of the first member 210 and the second member 220, as shown in FIG. 9, a rod-shaped jig 300 is used to forcibly force the elastically deformed portion 213 of the first member 210. The second member 220 may be pulled out from the first member 210 in the axial direction X in a state where the second member 220 is bent outward in the radial direction and deformed. Further, in order to release the assembly of the first member 210 and the second member 220 using this jig 300, the claw portion 214 of the elastically deformed portion 213 of the first member 210 becomes the groove portion 223 of the second member 220. A part of the inclined surface 215 is radially outside the outer surface of the second cylinder portion 221 of the second member 220 so that the jig 300 can come into contact with the inclined surface 215 of the claw portion 214 while being caught. It needs to be located. According to this configuration, the assembly of the first member 210 and the second member 220 can be released by using the jig 300.

By the way, in the above-mentioned third embodiment, the elastically deformed portion 213 of the first member 210 and the groove portion 223 of the second member 220 each correspond to the "locking portion" described in the claims.

Further, in the third embodiment described above, the first member 210 has an elastically deformed portion 213 provided with a claw portion 214 projecting radially inward from the plate-shaped main body, and the second member 220 is a second member. It has a groove portion 223 provided on the outer surface of the tubular portion 221 so as to be recessed inward in the radial direction. However, the present invention is not limited to this, and the first member 210 has a groove portion provided on the inner surface of the first tubular portion 211 so as to be recessed outward in the radial direction, and the second member 220 is provided. , It may have an elastically deformed portion provided with a claw portion protruding radially outward from the plate-shaped main body.

In the first, second, and third embodiments described above, the culture membrane 10 has a membrane body 11 and an annular support frame 12 joined to the peripheral edge of the membrane body 11. However, the present invention is not limited to this, and if the support frame 12 has a frame-like structure joined to the film body 11, for example, it is formed in a cross shape or a net shape when viewed from the axial direction X. You may be.

The present invention is not limited to the above-described embodiments and modifications, and various modifications can be made without departing from the spirit of the present invention.

1,100,200: Cell culture instrument, 10: Culture membrane, 11: Membrane body, 12: Support frame, 20,110,210: First member, 21,111,211: First cylinder part, 21a: Peripheral wall part , 21b: Notch, 22,212: Placement, 23: Side opening, 25,124: Recess, 25a: Axis parallel, 25b: Orthogonal, 30,120,220: Second member, 31,121 , 221: 2nd cylinder part, 32,122,222: Opposing part, 35,113: Convex part, 213: Elastic deformation part, 214: Claw part, 215: Inclined surface, 223: Groove part.

Claims (8)

A culture membrane having a membrane body and a support frame bonded to the membrane body,
A first having a first tubular portion extending in a tubular shape in the axial direction and a mounting portion on which the culture film is placed in contact with the support frame on which an axial end face facing the axial direction is formed. Members and
A second having a second tubular portion extending in a tubular shape in the axial direction and an opposing portion that faces the axial end surface of the previously described mounting portion in the axial direction and sandwiches the support frame between the previously described mounting portion. Members and
It is a cell culture device equipped with
The first member and the second member are locking portions that regulate relative movement in the axial direction in which the above-mentioned placing portion and the facing portion are assembled from each other so as to sandwich the support frame. A cell culture device that has. The locking portion is
A recess provided on one of the first cylinder portion and the second cylinder portion and extending in the circumferential direction,
A convex portion provided on the other side of the first tubular portion and the second tubular portion, which moves relative to the concave portion due to relative rotation of the first member and the second member in the circumferential direction and fits into the concave portion. ,
The cell culture apparatus according to claim 1. The recess is provided in the peripheral wall of one of the first cylinder portion and the second cylinder portion in a radial direction.
The cell culture device according to claim 2, wherein the convex portion is provided so as to project in the radial direction on the other peripheral wall of the first tubular portion and the second tubular portion. The locking portion is
An elastically deformable portion that is elastically deformable in the radial direction and is provided on one of the first cylinder portion and the second cylinder portion and has a claw portion that protrudes in the radial direction at the tip portion in the axial direction.
A groove portion provided on the other side of the first cylinder portion and the second cylinder portion on which the claw portion is caught when the first member and the second member approach a predetermined positional relationship in the axial direction.
The cell culture apparatus according to claim 1. The first member radially penetrates a predetermined portion including a portion on the peripheral wall of the first cylinder portion on the side of the previously described placement portion in the axial direction with respect to the culture membrane placed at least on the previously described placement portion. The cell culture apparatus according to any one of claims 1 to 4, which is provided in the above and has a side opening which opens a part of the first cylinder portion. The first tubular portion has a peripheral wall portion whose inner peripheral surface is formed so that the culture film can be guided to the previously described placement portion.
1. 5. The cell culture device according to any one of 5. The first tubular portion has a peripheral wall portion whose inner peripheral surface is formed so that the culture film can be guided to the previously described placement portion.
The peripheral wall portion has a notch portion that penetrates in the radial direction and is provided so as to extend in the axial direction from the axial end portion of the first tubular portion to the portion where the previously described placing portion is provided.
The cell culture apparatus according to claim 5, wherein the side opening and the notch are provided at the same position in the circumferential direction of the first member and are continuously arranged in the axial direction. The first tubular portion has a peripheral wall portion whose inner peripheral surface is formed so that the culture film can be guided to the previously described placement portion.
The peripheral wall portion has a notch portion that penetrates in the radial direction and is provided so as to extend in the axial direction from the axial end portion of the first tubular portion to the portion where the previously described placing portion is provided.
The cell culture apparatus according to claim 5, wherein the side opening and the notch are provided at different positions in the circumferential direction of the first member.

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