Culture Insert

Abstract

To provide a culture insert which does not cause a tip of a pipet to interfere with a well plate when the pipet is inserted into the well plate, and enables the tip of the pipet to be sufficiently inserted to the bottom portion of the well plate.SOLUTION: A culture insert 100 comprises a cylinder portion 11 and an insertion port 61 into which a pipe in a posture parallel with the axis 11L of the cylinder 11 can be inserted.SELECTED DRAWING: Figure 23

Description

[Technical field]

[0001] The present invention relates to a culture insert. [Background technology]

[0002] In cell culture, a culture insert with a membrane on the bottom on which cells are placed is inserted into a well such as a well plate or petri dish, which is a cell culture medium container, and the membrane is placed in the culture medium to culture the cells. In this case, a culture insert provided with a member known as a positioning arm has been disclosed for stabilizing the position of the culture insert within the well plate (Patent Document 1). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] JP 2020-110114 A Summary of the Invention [Problem to be solved by the invention]

[0004] In the above-mentioned cell culture process, when replacing the medium in the well, this may be done by inserting a pipette into the gap between the well and the culture insert while the culture insert remains placed inside the well. However, when a pipette is inserted into a well plate, if the tip of the pipette touches the insert, the membrane vibrates or shifts, which can cause cells to shift on the membrane or cause shear forces between the medium and the membrane, damaging the cells.

[0005] In addition, when changing the medium, it may not be possible to insert the pipette all the way to the bottom of the well plate. In this case, it is necessary to tilt the well plate with the culture insert still inserted, and shift the medium at the bottom of the well plate so that the tip of the pipette can reach the medium. This also causes damage to the cells as described above.

[0006] The present invention has been made in consideration of the above-mentioned circumstances, and has an object to provide a culture insert that prevents the tip of a pipette from interfering with insertion of the pipette into a well plate, and allows the tip of the pipette to be inserted sufficiently to the bottom of the well plate. [Means for solving the problem]

[0007] In order to solve the above problems, the present invention proposes the following means. The culture insert according to the present invention comprises a cylindrical portion and an insertion opening into which a pipe parallel to the axis of the cylindrical portion can be inserted.

[0008] According to this invention, the well plate is provided with an insertion port into which a pipe parallel to the axis of the cylindrical portion can be inserted. This allows the tip of the pipe to be inserted into the well plate through the insertion port. In other words, it is possible to prevent interference between the tip of the pipe and the culture insert when inserting the pipe into the well plate. This prevents the culture insert inserted into the well plate from vibrating or shifting.

[0009] Furthermore, the tip of the pipe can be inserted sufficiently deep into the bottom of the well plate, which allows the tip of the pipe to reach the medium without tilting the well plate when replacing the medium using the pipe. These can contribute to more stable cell culture.

[0010] The nozzle may further include a flange provided on the cylindrical portion.

[0011] According to this invention, the cylindrical portion is provided with a flange portion, whereby when the cylindrical portion is inserted into the well plate, the position of the culture insert can be stabilized by bringing the flange portion into contact with the well plate.

[0012] An opening may be provided between the flange portion and the cylindrical portion.

[0013] According to this invention, an opening is provided between the flange and the cylindrical part. This allows, for example, a region through which air can pass between the inner wall of the well plate and the cylindrical part. This prevents air pressure from acting as a resistance when inserting or removing the culture insert, and allows the culture insert to be inserted or removed more stably.

[0014] The culture insert may also be provided with a mark for image recognition that is visible when viewed in plan.

[0015] According to this invention, a mark for image recognition that is visible in a plan view of the culture insert is provided, which makes it easier for the machine to identify the position of the insertion port when the culture medium exchange using the pipe is automated by the machine.

[0016] The device may also include legs provided on the cylindrical portion.

[0017] According to this invention, the tube portion is provided with legs, which allows the culture insert to be entirely contained within the well plate, thereby preventing the culture insert from being affected by the surrounding space, for example, when there is limited space around the well plate.

[0018] The insertion opening may be provided on an outer wall of the cylindrical portion.

[0019] According to this invention, the insertion opening is provided on the outer wall of the tubular portion, which makes it possible to provide the insertion opening without adding a special member to the culture insert.

[0020] Also, a plurality of the insertion openings may be provided.

[0021] According to the present invention, a plurality of insertion openings are provided, which allows for more flexible response to cases where the surrounding space is limited, such as when inserting a pipe or when changing the culture medium by machine.

[0022] Also, a plurality of the insertion openings having different sizes may be provided.

[0023] According to this invention, a plurality of insertion openings of different sizes are provided, which allows the size of the pipe to be selected as needed, thereby contributing to more efficient culture medium exchange.

[0024] A wall portion having an end connected to an inner wall of the cylindrical portion may be provided inside the cylindrical portion, and the insertion opening may be provided in the wall portion.

[0025] According to this invention, the insertion port is provided in a wall portion provided on the inner wall of the cylindrical portion. This allows the pipe to be inserted from inside the cylindrical portion toward the well plate. In other words, the pipe can be inserted from a portion closer to the center of the well plate. This allows for more efficient medium exchange compared to inserting the pipe from the end of the well plate. Effect of the Invention

[0026] According to the present invention, it is possible to provide a culture insert that allows the tip of a pipette to be inserted sufficiently to the bottom of a well plate without interference with the tip of the pipette when the pipette is inserted into the well plate. [Brief description of the drawings]

[0027] [Figure 1] FIG. 2 is a plan view of a first example of a culture insert according to the present invention. [Diagram 2]FIG. 2 is a side view of the culture insert shown in FIG. 1 in the direction II. [Diagram 3] FIG. 2 is a perspective view of the culture insert shown in FIG. 1. [Figure 4] FIG. 2 is a plan view of a second example of a culture insert according to the present invention. [Diagram 5] FIG. 5 is a side view of the culture insert shown in FIG. 4 in the V direction. [Figure 6] FIG. 5 is a perspective view of the culture insert shown in FIG. [Figure 7] FIG. 2 is a plan view of a third example of a culture insert according to the present invention. [Figure 8] FIG. 8 is a side view of the culture insert shown in FIG. 7 in the VIII direction. [Figure 9] FIG. 8 is a perspective view of the culture insert shown in FIG. [Figure 10] FIG. 11 is a plan view of a fourth example of a culture insert according to the present invention. [Figure 11] 11 is a side view of the culture insert shown in FIG. 10 in the XI direction. [Figure 12] FIG. 11 is a perspective view of the culture insert shown in FIG. [Figure 13] FIG. 2 is a plan view of a first example of a leg-type culture insert according to the present invention. [Figure 14] FIG. 14 is a side view of the leg-type culture insert shown in FIG. 13 in the XIV direction. [Figure 15] FIG. 14 is a perspective view of the leg-type culture insert shown in FIG. [Figure 16] FIG. 2 is a plan view of a second example of a leg-type culture insert according to the present invention. [Figure 17] 17 is a side view of the leg-type culture insert shown in FIG. 16 in the XVII direction. [Figure 18] FIG. 17 is a perspective view of the leg-type culture insert shown in FIG. 16. [Figure 19] FIG. 11 is a plan view of a third example of a leg-type culture insert according to the present invention. [Figure 20] FIG. 20 is a side view of the leg-type culture insert shown in FIG. 19 in the direction XX. [Figure 21] FIG. 20 is a perspective view of the leg-type culture insert shown in FIG. 19. [Figure 22] This is a modified example of the leg-type culture insert shown in FIG. [Figure 23] FIG. 2 is a schematic cross-sectional view of the culture insert according to the present invention attached to a well plate. [Figure 24] FIG. 2 is a schematic cross-sectional view of the stem-type culture insert according to the present invention attached to a well plate. [Diagram 25] FIG. 2 is a diagram showing a state in which a pipette is inserted into the insertion port of the culture insert according to the present invention. [Figure 26] FIG. 2 is a diagram showing a state in which a pipette is inserted into the insertion port of the stem-type culture insert according to the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] Hereinafter, a culture insert 100 according to one embodiment of the present invention will be described with reference to the drawings. As shown in FIGS. 1, 2, and 3, the culture insert 100 includes a cylindrical portion 11, a flange portion 21, a tapered portion 31, an opening 41, a marker 51, an insertion port 61, and a membrane 71. 23, the culture insert 100 is used by inserting one end of the tubular portion 11 into a well W of a well plate WP with one end facing up and the other end facing down. Specifically, the membrane 71 of the culture insert 100 is inserted so as to be positioned inside the culture medium C placed at the bottom of the well W (details will be described later). In this embodiment, the culture insert 100 is preferably manufactured by injection molding using polycarbonate.

[0029] The well plate WP is a plate having a plurality of wells W. The well plate WP accommodates a culture insert 100 inside each well W. The wells W are depressions provided in the well plate WP. The medium C for cell culture is placed at the bottom of the depression. The wells W provided in the well plate WP may be connected to each other. The culture insert 100 is inserted so that the membrane 71 is disposed inside the culture medium C disposed in the well W. In this embodiment, a well plate WP that is well known in the art is preferably used. The size of the culture insert 100 is appropriately determined according to the size of the well W of the well plate WP to be used.

[0030] The tubular portion 11 is a portion that is placed inside the well W of the well plate WP. In this embodiment, the tubular portion 11 is cylindrical. As shown in FIG. 23, when the tubular portion 11 is inserted into the well W, the tubular portion axis 11L (axis), which is the central axis of the tubular portion 11, is parallel to the well axis WL, which is the central axis of the well W. The outer diameter of the cylinder is set according to the inner diameter of the well W into which it is to be inserted. For example, when the inner diameter of the well W is ** mm, it is preferable that the outer diameter of the tubular portion 11 be ** mm to ** mm and the inner diameter be ** mm to ** mm.

[0031] The flange 21 is an annular flange provided on one end of the tube 11. The outer diameter of the flange 21 is preferably larger than the inner diameter of the well W, and the inner diameter of the flange 21 is preferably smaller than the inner diameter of the well W. As shown in FIG. 2, the flange 21 is provided with a step 21s. As a result, as shown in FIG. 23, when the culture insert 100 is inserted into the well W, the portion of the step 21s facing the upper surface 31a of the well plate WP contacts the upper surface 31a of the well plate WP, and the portion of the step 21s facing the inner wall of the well W contacts the inner wall of the well W or has a minute gap. As a result, when the culture insert 100 is inserted into the well W, the other end of the tube 11 is prevented from directly contacting the bottom of the well W, and the culture insert 100 has a role of holding the culture insert 100 in the well W so that it does not move.

[0032] The tapered portion 31 connects between the tubular portion 11 and the flange portion 21. As shown in FIG. 3, the tapered portion 31 has an upper surface 31a and an inner surface 31b. The inner diameter of the inner surface 31b decreases from the flange portion side to the tubular portion side. This shape connects the flange portion 21 and the tubular portion 11, which has a smaller diameter than the flange portion 21. The tapered portion 31 is provided with an opening 41 and a marker 51.

[0033] The opening 41 is formed by cutting out a part of the tapered portion 31 provided between the flange portion 21 and the cylindrical portion 11. This ensures an area in which the opening 41 is formed at one end of the cylindrical portion 11. In other words, the tapered portion 31 is provided with a plurality of tapered pieces arranged at intervals in the circumferential direction, and the opening 41 is formed between the tapered pieces adjacent to each other in the circumferential direction. 10, 11, and 12, the opening 41 may be provided continuously from the flange 21 to the outer wall of the tubular part 11. In this case, an area through which air can pass can be provided between the inner wall of the well W and the tubular part 11. This prevents air pressure from acting as resistance when inserting or removing the culture insert 100, making it possible to insert or remove the culture insert 100 more stably.

[0034] The mark 51 is provided on the upper surface 31a of the tapered portion 31. That is, in a plan view of the culture insert 100, as shown in Figs. 2 and 3, the mark 51 is provided by molding a figure such as a circle, triangle, or square into a convex or concave shape so as to be visible. Alternatively, the mark 51 may be provided by applying paint. By using the positional relationship between the mark 51 and the insertion opening 61 for image recognition, for example, when the culture medium is replaced by a machine, the machine recognizes the mark 51 and determines the position of the insertion opening 61.

[0035] The insertion port 61 is a portion for inserting a pipette P (pipe) when performing culture medium replacement with the culture insert 100 inserted in the well plate WP. The insertion port 61 includes an opening 41, a first recess 61a formed in the tubular portion 11, and a second recess 61b formed in the flange portion 21.

[0036] As described above, the opening 41 is formed by cutting out a part of the tapered portion 31. As a result, when the culture insert 100 is viewed from one end side, as shown in Fig. 2, a part of one end of the tubular portion 11 is released. The first recess 61a, the second recess 61b, and the opening 41 are positioned at the same circumferential position.

[0037] The first recess 61a is formed to be recessed from the outside to the inside of the tubular portion 11. As shown in FIG. 2, the insertion opening 61 is tapered. An insertion opening axis 61L, which is the central axis of the tapered shape of the first recess 61a, is parallel to the tubular portion axis 11L of the tubular portion 11. This ensures that when the pipette P is inserted, the pipette axis PL, which is the central axis of the pipette P, is inserted in a state parallel to the tubular portion axis 11L of the tubular portion 11 and the well axis WL of the well W.

[0038] The second recess 61b is formed by cutting out the collar 21 in an arc shape from the inner periphery to the outer periphery of the annular collar 21. As a result, as shown in Fig. 2, the arcs forming the first recess 61a and the second recess 61b are concentric when the culture insert 100 is viewed from the first end side. This prevents the pipette P from interfering with the collar 21 and the cylindrical portion 11 when the pipette P is inserted from the first end side of the cylindrical portion 11. In this embodiment, a known, general pipette P is preferably used. A special pipe or the like may be used depending on a special purpose. The tip of the pipette P has a tapered shape. The first recess 61a supports a portion of the pipette P closer to the lower end than the second recess 61b from the side.

[0039] A plurality of insertion openings 61 may be provided in the tubular portion 11. In this case, as shown in Fig. 4, Fig. 5, and Fig. 6, the insertion openings 61 may be different in size (for example, 61s, 61m, 61l), or a plurality of the same size may be provided. In this case, the first recess 61a and the second recess 61b may also vary in size (for example, 61as, 61am, 61al and 61bs, 61bm, 61bl). 7, 8 and 9, a wall portion 11W whose end is connected to the inner wall of the tube portion 11 may be provided, and the insertion opening 61 may be provided in this wall portion 11W. In this case, the second recess 61b is not formed in the flange portion 21, and the insertion opening 61 is formed only by the first recess 61a.

[0040] The membrane 71 is provided at the other end of the tube portion 11 so as to close the opening 41 on the inside of the tube portion 11. The membrane 71 is, for example, a known semipermeable membrane. When cell culture is performed using the culture insert 100, cells are placed on the membrane 71.

[0041] Next, the leg-type culture insert 200 according to the present invention will be described. Note that for the leg-type culture insert 200, the same components as those in the culture insert 100 are given the same reference numerals, and their explanation will be omitted, and only the differences will be described. As shown in FIGS. 13, 14 and 15, the leg-type culture insert 200 includes a tube portion 11, an insertion opening 61, a leg portion 22 and a membrane 71.

[0042] The leg portion 22 is a portion provided at the other end of the tubular portion 11. As shown in FIG. 15, a plurality of leg portions 22 are provided at intervals at the other end of the tubular portion 11. As shown in FIG. 24, when the leg-type culture insert 200 is inserted into the well W, the leg portion 22 comes into contact with the bottom of the well W. This allows the leg-type culture insert 200 to stand independently on the bottom of the well W. This serves to position the leg-type culture insert 200 in the well W without allowing the flange portion 21 to protrude outside the well W and without the membrane 71 coming into contact with the bottom of the well W.

[0043] In the leg-type culture insert 200, the insertion opening 61 provided in the tubular portion 11 can have the same structure as the culture insert 100. For example, as shown in Figures 16, 17, and 18, insertion openings 61 of different sizes (e.g., 61s, 61mM, 61l) may be provided, or multiple openings of the same size may be provided. Note that the insertion opening 61 in the leg-type culture insert 200 does not have a flange portion 21 in which the second recess 61b is provided, and is therefore formed only by the first recess 61a. 19, 20 and 21, a wall portion 11W to be connected to the end portion of the inner wall of the tube portion 11 may be provided, and the insertion opening 61 may be provided in this wall portion 11W.

[0044] The culture insert 100 and the leg-type culture insert 200 according to this embodiment are mainly used for cell culture. That is, with cells arranged on the membrane 71, the culture insert 100 or the leg-type culture insert 200 is inserted into the inside of the well W of a well plate WP, which is a cell culture medium container, as shown in Figs. 23 and 24. As a result, the membrane 71 on which the cells are arranged is placed in the medium C in the well plate WP. As a result, the medium C that has permeated the membrane 71 is supplied to the cells arranged on the membrane 71. Cell culture is performed by maintaining this state.

[0045] During cell culture under these conditions, medium C may become contaminated by waste products released from the cultured cells, the nutrients contained in medium C may become depleted, or the components of medium C may change due to oxidation, so it is necessary to replace medium C from time to time. At this time, as shown in Figures 23 and 24, the insertion port axis 61L of the insertion port 61 is parallel to the cylindrical portion axis 11L of the cylindrical portion 11. The cylindrical portion axis 11L of the cylindrical portion 11 is also parallel to the well axis WL of the well W of the well plate WP into which the culture insert 100 and the foot-type culture insert 200 are inserted. That is, when changing the culture medium, as shown in Figures 25 and 26, the pipette P can be inserted into the well plate WP with the pipette axis PL of the pipette P parallel to the cylindrical portion axis 11L of the cylindrical portion 11, that is, with the well axis WL of the well W and the pipette axis PL of the pipette P parallel to each other. This allows the tip of the pipette P to be efficiently inserted into the bottom of the well W.

[0046] As described above, the culture insert 100 and the leg-type culture insert 200 according to this embodiment are provided with an insertion port 61 into which a pipette P can be inserted in an orientation parallel to the cylindrical axis 11L of the cylindrical portion 11. This allows the tip of the pipette P to be inserted into the well plate WP through the insertion port 61. In other words, it is possible to prevent interference between the tip of the pipette P and the culture insert 100 and the leg-type culture insert 200 when inserting the pipette P into the well plate WP. This makes it possible to prevent the culture insert 100 and the leg-type culture insert 200 inserted into the well plate WP from vibrating or shifting.

[0047] Furthermore, the tip of the pipette P can be fully inserted to the bottom of the well plate WP. This allows the tip of the pipette P to reach the medium C without tilting the well plate WP when replacing the medium C with the pipette P. These can contribute to more stable cell culture.

[0048] Furthermore, the cylindrical portion 11 is provided with a flange portion 21. With this, when the cylindrical portion 11 is inserted into the well plate WP, the position of the culture insert 100 can be stabilized by bringing the flange portion 21 into contact with the well plate WP.

[0049] Also, an opening 41 is provided between the flange 21 and the tubular portion 11. This makes it possible to provide an area through which air can pass, for example, between the inner wall of the well plate WP and the tubular portion 11. This prevents air pressure from acting as resistance when inserting or removing the culture insert 100, making it possible to insert or remove the culture insert 100 more stably.

[0050] Also, a mark 51 for image recognition is provided which is visible in a plan view of the culture insert 100. This makes it easier for the machine to identify the position of the insertion port 61 when automating the medium exchange using the pipette P.

[0051] Furthermore, the cylindrical portion 11 is provided with legs 22. This allows the entire leg-type culture insert 200 to be stored inside the well plate WP. Therefore, for example, when there is limited space around the well plate WP, it is possible to prevent the culture insert 200 from being affected by the surrounding space.

[0052] Furthermore, the insertion opening 61 is provided on the outer wall of the tubular portion 11. This allows the insertion opening 61 to be provided in the culture insert 100 and the leg-type culture insert 200 without adding any special member.

[0053] In addition, a plurality of insertion openings 61 are provided. This allows for more flexible response when inserting the pipette P, when changing the culture medium mechanically, or when there is a restriction on the surrounding space.

[0054] In addition, a plurality of insertion openings 61 of different sizes are provided, which allows pipettes P of different sizes to be used as needed, thereby contributing to more efficient culture medium exchange.

[0055] Also, an insertion port 61 is provided in a wall portion 11W provided on the inner wall of the tube portion 11. This allows the pipette P to be inserted from the inside of the tube portion 11 toward the well plate WP. In other words, the pipette P can be inserted into the insertion port 61 from a position closer to the center of the well plate WP. This allows for more efficient medium exchange compared to the case where the pipette P is inserted from the end of the well plate WP.

[0056] The technical scope of the present invention is not limited to the above-described embodiment, and various modifications can be made without departing from the spirit of the present invention. For example, the tube portion 11 may be in a rectangular tube shape instead of a cylindrical shape. Furthermore, the flange portion 21 does not have to be annular, and may be provided only at a portion corresponding to the upper surface 31a of the tapered portion 31. In other words, the flange portion 21 may be open to match the insertion opening 61. Furthermore, the flange portion 21 and the leg portion 22 may be provided simultaneously on one culture insert 100 . As shown in FIG. 22, the leg-type culture insert 200 may be provided with a knob 11h. Furthermore, the culture insert 100 and the leg-type culture insert 200 according to this embodiment are not limited to being used in the well W of the well plate WP, but may also be used in a single petri dish, petri dish, or the like.

[0057] In addition, within the scope of the invention, the components in the above-described embodiments may be replaced with well-known components, and the above-described modified examples may be combined as appropriate. [Explanation of symbols]

[0058] 11 Cylinder part 11L axis 11W wall 21 Flange 22 Legs 41 Opening 51 signs 61 Insertion port 100 Culture Inserts

Claims

1. A cylindrical portion and an insertion port into which a pipe parallel to an axis of the cylindrical portion can be inserted; Equipped with The insertion opening includes a first recess formed to be recessed from the outside to the inside of the cylindrical portion. Culture insert.

2. A flange portion is provided on the cylindrical portion. The culture insert according to claim 1 .

3. An opening is provided between the flange portion and the cylindrical portion. The culture insert according to claim 2.

4. A mark for image recognition visible in a plan view of the culture insert is provided; A culture insert according to claim 2 or 3.

5. The cylindrical portion has a leg portion provided thereon. The culture insert according to claim 1 .

6. The insertion port is provided on an outer wall of the cylindrical portion. A culture insert according to any one of claims 1 to 5.

7. A plurality of the insertion openings are provided. A culture insert according to any one of claims 1 to 6.

8. A plurality of the insertion openings having different sizes are provided. A culture insert according to any one of claims 1 to 7.

9. A wall portion is provided inside the cylindrical portion, and an end portion of the wall portion is connected to an inner wall of the cylindrical portion, The insertion opening is provided in the wall portion. A culture insert according to any one of claims 1 to 8.

10. A cylindrical portion, an insertion port into which a pipe parallel to an axis of the cylindrical portion can be inserted; Equipped with The insertion port is provided in a wall portion whose end is connected to the inner wall of the cylindrical portion. Culture insert.

Images