Cell culture kits

Abstract

To provide a novel cell culture kit that allows easy manipulation of well contents.SOLUTION: A cell culture kit comprises a well plate having a plurality of wells, and a cell culture insert to be inserted into the wells. The cell culture insert has a cylindrical main body having a bottom, and a support part extending radially outward of the main body at the top of the main body. The support part is provided at a position to expose at least a part of the well in a posture of the cell culture insert being inserted into a fixed position in the well. The well has a sampling part in which the distance between the outer surface of the main body and the inner wall of the well is relatively wide in the circumferential direction of the well when in that posture.SELECTED DRAWING: Figure 2

Description

[Technical field]

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

[0002] Conventionally, a method of culturing cells by combining a well plate and a cell culture insert has been known (see, for example, Patent Document 1). Hereinafter, the cell culture insert may be simply referred to as an "insert." The combination of the well plate and the insert may be referred to as a "cell culture kit."

[0003] An insert is a cylindrical culture device with a bottom plate used for culturing cells, organs, and tissues, and is inserted into the well of a well plate. Usually, the bottom of the insert is made of a membrane that is permeable to the culture medium stored in the well. By seeding cells, etc. on the inner bottom of the insert and inserting it into the well, the culture medium in the well can be distributed to the cells, etc. through the bottom (membrane), allowing the cells, etc. to be cultured. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2020-110114 A Summary of the Invention [Problem to be solved by the invention]

[0005] During cell culture, the contents of the wells (e.g., culture medium) may need to be manipulated for component analysis or replacement. However, with conventional cell culture kits, it is difficult to manipulate the contents of the wells, and improvements were needed.

[0006] The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a novel cell culture kit that allows easy manipulation of the contents of the wells. [Means for solving the problem]

[0007] One possible scenario in which a problem occurs with cell culture kits is when, during cell culture, a tool such as a pipette is used to remove the medium from the well in order to analyze or replace the medium. In this case, it is conceivable to remove the medium from the well after removing the insert from the well. However, in this case, there is a risk that the cells removed from the medium in the well will dry out or be exposed to air, which may reduce the cell viability.

[0008] Furthermore, if the above operation is performed with the insert inserted in the well, it is necessary to insert a tool such as a pipette into the gap between the well and the insert. However, in such a cell culture kit, the size of the insert is limited in order to form a gap through which the tool can be inserted, and the efficiency of cell culture is reduced. Therefore, there is still room for improvement in conventional cell culture kits.

[0009] The inventors have conducted extensive research into a cell culture kit capable of solving these problems and have completed the invention.

[0010] That is, in order to solve the above problems, one aspect of the present invention includes the following aspects.

[0011] [1] A cell culture kit comprising: a well plate having a plurality of wells; and a cell culture insert to be inserted into the wells, the cell culture insert having a cylindrical main body portion having a bottom, and a support portion extending radially outward from an upper portion of the main body portion, the support portion being positioned to expose at least a portion of the well when the cell culture insert is inserted into a fixed position in the well, the well having a sampling portion in which the distance between an outer surface of the main body portion and an inner wall of the well is relatively wide in the circumferential direction of the well when in the fixed position.

[0012] [2] The cell culture kit described in [1], wherein the multiple wells are arranged in a matrix in a planar view, and each of the multiple wells has the sampling portion facing in the same direction in a planar view.

[0013] [3] A cell culture kit as described in [1] or [2], wherein the main body portion is cylindrical, the well has a corner that protrudes outside an imaginary circle inscribed in the well, and the sampling portion is a gap formed between the main body portion and the corner of the well in the attitude.

[0014] [4] The cell culture kit according to [3], wherein the well has a rectangular shape when viewed in plan.

[0015] [5] A cell culture kit described in any one of [1] to [4], wherein the cell culture insert has a plurality of support portions discretely arranged in the circumferential direction of the main body portion, and the upper end portion of the well has an anti-rotation portion that holds the support portions and determines the circumferential posture of the cell culture insert.

[0016] [6] A cell culture kit described in any one of [1] to [5], wherein the cell culture insert has a protrusion extending radially from the outer surface of the main body portion, and the protrusion interferes with the inner wall of the well in the said position. Effect of the Invention

[0017] According to the present invention, a novel cell culture kit can be provided that allows easy manipulation of the contents of the wells. [Brief description of the drawings]

[0018] [Figure 1] FIG. 1 is a schematic perspective view showing a cell culture kit 100 of the first embodiment. [Diagram 2] FIG. 2 is a partially enlarged view of the cell culture kit 100. As shown in FIG. [Diagram 3] FIG. 3 is a schematic cross-sectional view of the cell culture kit 100. [Figure 4]FIG. 4 is a partially enlarged view of the cell culture kit 100. [Diagram 5] FIG. 5 is an explanatory diagram for explaining problems that may occur due to the presence of the sampling unit 135. [Figure 6] FIG. 6 is a schematic perspective view showing a cell culture kit 200 according to the second embodiment. [Figure 7] FIG. 7 is a partially enlarged view of the cell culture kit 200. As shown in FIG. [Figure 8] FIG. 8 is a schematic cross-sectional view of a cell culture kit 200. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] [First embodiment] Hereinafter, a cell culture kit according to a first embodiment of the present invention will be described with reference to Figures 1 to 5. In all the following drawings, the dimensions and ratios of each component are appropriately changed to make the drawings easier to see.

[0020] In the following description, an xyz Cartesian coordinate system is set, and the positional relationship of each member is described with reference to this xyz Cartesian coordinate system. Here, a specific direction in a horizontal plane is defined as the x-axis direction, a direction perpendicular to the x-axis direction in the horizontal plane is defined as the y-axis direction, and a direction perpendicular to both the x-axis direction and the y-axis direction (i.e., the vertical direction) is defined as the z-axis direction.

[0021] In the following description, the term "planar view" refers to a field of view in which an object is observed from the +z direction.

[0022] Fig. 1 is a schematic perspective view showing a cell culture kit 100 of the present embodiment. Fig. 2 is a partially enlarged view of the cell culture kit 100. Fig. 3 is a schematic cross-sectional view of the cell culture kit 100, taken along line III-III in Fig. 1.

[0023] As shown in FIGS. 1 to 3, the cell culture kit 100 includes a well plate 10 and a cell culture insert 20 (insert 20).

[0024] (Well plate) The well plate 10 has a top plate 11, a frame 12 that supports the top plate 11, and a plurality of wells 13 (six in the figure) provided in the top plate 11.

[0025] The top plate 11 has a rectangular shape in a plan view, and is connected to and supported by an inner peripheral surface 12a of a cylindrical frame body 12 that also has a rectangular shape in a plan view.

[0026] The frame 12 has a first frame 121 that is rectangular in plan view, and a second frame 122 that is continuous with the lower end of the first frame 121 on the outside of the first frame 121. An upper end 122a of the second frame 122 functions as a support portion that supports a lid (not shown) when the well plate 10 is covered with the lid.

[0027] An upper end 12b of the frame 12 is located above an upper surface 11a of the top plate 11. When the well plate 10 is covered with a lid (not shown), the upper end 12b functions as a support portion that supports the lid.

[0028] Wells 13 store a medium used for cell culture. Wells 13 are arranged in a matrix of 2 rows and 3 columns on top plate 11. Two sampling portions 135 are formed in wells 13. As shown in Fig. 1, each of the multiple wells 13 has sampling portions 135 facing in the same direction in a plan view. The shape of the well 13 and the sampling portion 135 will be described in detail later.

[0029] The bottom of the well 13 may have various known shapes. The bottom of the well 13 may be a flat bottom, a round bottom, or a V-bottom.

[0030] A plurality of (three in the figure) ridges 15 are provided along the edge of opening 13a in plan view at the upper end of well 13. ridges 15 correspond to the "anti-rotation portion" of the present invention.

[0031] Three recesses 15a are formed between two adjacent convex ridges 15 in the circumferential direction of the opening 13a. In FIG. 2, the lowest position of the recess 15a is shown to be at the same height as the upper surface 11a of the top plate 11, but this is not limited thereto and may be higher than the upper surface 11a. A plurality of recesses 15a are provided discretely in the circumferential direction of the opening 13a. The well plate 10 has a total of three recesses 15a at equal intervals of 120° in the circumferential direction of the opening 13a.

[0032] The well plate 10 is made of a material known as a material for well plates, for example, polystyrene.

[0033] (Cell Culture Insert) The insert 20 has a body portion 21 and a support portion 22 .

[0034] The main body 21 has a bottom 21a at the lower end (-z side) and is cylindrical with an open upper end (+z side). The internal space 21x of the main body 21 is used as a space for holding and culturing the target cells. The bottom 21a is made of a membrane that is permeable to the culture medium stored in the well 13.

[0035] The main body 21 is preferably cylindrical, but may have various shapes as long as it has a bottom plate and is tubular. The "cylindrical" main body 21 may have parallel outer surfaces in a cross section taken along the central axis of the main body 21, or may have a truncated cone shape in which the outer surfaces are not parallel in the cross section and the outer diameter decreases from the upper end to the lower end.

[0036] The support portion 22 is provided on an upper portion of the outer surface 21b of the main body portion 21. A plurality of support portions 22 are provided discretely in the circumferential direction of the main body portion 21. The insert 20 has a total of three support portions 22, equally spaced at intervals of 120° in the circumferential direction of the main body portion 21. Each support portion 22 extends radially outward from the main body portion 21.

[0037] 2, when the insert 20 is inserted into the well 13, the support parts 22 contact the upper surface 11a of the top plate 11 in each of the three recesses 15a. That is, the three support parts 22 are disposed in the three recesses 15a, respectively.

[0038] As a result, the support portion 22 supports the insert 20 with the bottom portion 21 a spaced apart from the bottom surface 13 b of the well 13 .

[0039] Furthermore, when the insert 20 is inserted into the well 13, the ridges 15 hold the support portions 22 in depressions 15a between adjacent ridges 15. Even if the insert 20 attempts to rotate in the circumferential direction of the body portion 21 within the well 13, the support portions 22 come into contact with the ridges 15. Therefore, the ridges 15 function as a rotation stopper that regulates the position of the insert 20 in the circumferential direction.

[0040] The insert 20 is made of a material known as a material for cell culture inserts, for example, polystyrene.

[0041] FIG. 4 is a partially enlarged view of the cell culture kit 100, and is a plan view showing the state in which the insert 20 is inserted into the well 13. As shown in FIG.

[0042] It should be noted that the insert 20 is inserted in a position expected in normal use when culturing cells in the well 13 using the cell culture insert. The "position expected in normal use" is the center of the well 13 in a plan view, and means the "fixed position of the well" when insert 20 is inserted into the well 13 in the present invention. In this sense, the "fixed position of the well" does not include a position in the well 13 where the insert 20 is excessively close to a portion of the inner wall of the well 13, or a position where the outer surface 21b of the insert 20 is in contact with the inner wall of the well 13.

[0043] As shown in Fig. 4, the shape of well 13 in plan view has corner 13x formed to protrude outward from imaginary circle VC inscribed in well 13. In detail, the shape of well 13 in plan view is a shape in which one side 131x of rectangle 131 is joined to semicircle 132 having side 131x as a chord. In Fig. 4, rectangle 131 is arranged on the +x side, and semicircle 132 is arranged on the -x side.

[0044] When the insert 20 having the cylindrical main body 21 is inserted into a fixed position of the well 13 having such a shape, in the inserted position, a part of the well 13 is exposed between the multiple support parts 22. That is, the support parts 22 are provided at positions that expose at least a part of the well 13.

[0045] Furthermore, in the position in which the insert 20 is inserted, the well 13 has a portion where the distance between the outer surface 21b of the main body 21 of the insert 20 and the inner wall 13c of the well 13 is relatively wider in the circumferential direction of the well 13. In a plan view, the distance W1 between the outer surface 21b of the main body 21 and a corner of the well 13 (a corner of the rectangle 131) is wider than the distance W2 between the outer surface 21b and an arc of the well 13 (an arc of a semicircle 132).

[0046] In FIG. 4, the "point where the distance between the outer surface 21b and the inner wall 13c of the well 13 is relatively wide" is indicated by reference numeral 135, and the "point between the outer surface 21b and the arc of the well 13" is indicated by reference numeral 135x. The point indicated by reference numeral 135 corresponds to the "sampling portion" in this invention. The well 13 in FIG. 4 has two sampling portions 135 corresponding to the corners of the rectangle 131. The sampling portions 135 are gaps formed between the main body portion 21 and the corners of the well 13 when the insert 20 is inserted at a fixed position in the well 13.

[0047] As described above, in sampling section 135, the distance between outer surface 21b and inner wall 13c of well 13 is relatively greater than in the portion indicated by reference symbol 135x. Therefore, in sampling section 135, it is easier to insert an instrument such as a pipette than in the portion indicated by reference symbol 135x, and the contents in well 13 can be easily manipulated.

[0048] As described above, the sampling parts 135 face the same direction in each of the multiple wells 13. Therefore, an operator using the cell culture kit 100 can operate the contents in each well 13 from the sampling part 135 of the well 13 without changing his / her posture with respect to the cell culture kit 100. Therefore, the cell culture kit 100 can improve the work efficiency.

[0049] 5 is an explanatory diagram illustrating a possible problem caused by the presence of the sampling part 135. When the well plate 10 has the sampling part 135, if the length of the support part 22 is shorter than the distance W1, it is possible that the support part 22 will fall into the well 13 in a position where the support part 22 and the sampling part 135 overlap in a plan view (position indicated by symbol X), causing the insert 20 to tilt. This situation may occur, for example, when the insert 20 rotates in the circumferential direction indicated by arrow α during operation. The length of the support part 22 means "the length from the outer surface 21b to the tip of the support part 22 in a plan view."

[0050] In contrast, in the cell culture kit 100, to address such anticipated problems, a convex ridge 15 is provided on the edge of the well 13 to restrict circumferential rotation of the insert 20. Therefore, in the cell culture kit 100, the position of the insert 20 inserted into the well 13 can be stabilized, and the problem of the insert 20 falling into the well 13 can be suppressed.

[0051] The cell culture kit 100 having the above-mentioned configuration is a novel cell culture kit that allows easy manipulation of the contents of the wells 13.

[0052] In the cell culture kit of the present embodiment, the well plate 10 has the convex ridge portion 15, but this is not limited thereto. Even if the well plate 10 does not have the convex ridge portion 15, the problem described in FIG. 5 can be suppressed by taking measures such as making the support portion 22 longer than the distance W1 or increasing the number of the support portions 22.

[0053] In the cell culture kit of the present embodiment, the sampling parts 135 provided in the multiple wells 13 face in the same direction, but this is not limited to this. Even if the sampling parts 135 of the wells 13 face in different directions, the effects of the present invention can be achieved.

[0054] In the well plate 10 of the cell culture kit of this embodiment, the well 13 has two sampling parts 135, but the number of sampling parts 135 can be set arbitrarily as long as it does not impair the effects of the present invention. The well plate 10 may have only one sampling part 135, or may have three or more sampling parts 135.

[0055] In the cell culture kit of this embodiment, the sampling part 135 is a corner, but is not limited to this. It may have another shape as long as it is formed as a part where the distance between the outer surface of the main body part and the inner wall of the well is relatively wide in the circumferential direction of the well and it is easier to insert an instrument into the well than other parts.

[0056] In the cell culture kit of the present embodiment, the insert 20 has a plurality of support parts 22, but this is not limited thereto. The support part 22 may have various shapes as long as it supports the insert 20 in the well 13 and does not impede the operation in the sampling part 135.

[0057] [Second embodiment] 6 to 8 are explanatory diagrams of a cell culture kit 200 according to a second embodiment of the present invention. In this embodiment, components common to those in the first embodiment are given the same reference numerals, and detailed descriptions thereof will be omitted.

[0058] Fig. 6 is a schematic perspective view showing the cell culture kit 200, and corresponds to Fig. 1. Fig. 7 is a partially enlarged view of the cell culture kit 200, and corresponds to Fig. 2. As shown in FIG. 6, the cell culture kit 200 includes a well plate 30 and a cell culture insert 40 (insert 40).

[0059] The well plate 30 has a top plate 31, a frame 32 that supports the top plate 31, and a plurality of wells 33 (six in the figure) provided in the top plate 31.

[0060] The top plate 31 and the frame 32 can have the same configuration as the top plate 11 and the frame 32 in the well plate 10 of the first embodiment.

[0061] The well 33 is rectangular in plan view, more specifically, square in plan view. The well 33 has four corners in plan view.

[0062] At the upper end of the well 33, a closed ring-shaped protruding portion 35 is provided along the edge of the opening in a plan view.

[0063] The insert 40 has a main body portion 41 and a support portion 42. The main body portion 41 can have a similar configuration to the main body portion 21 of the insert 20 of the first embodiment.

[0064] The support portion 42 is provided on an upper portion of the outer surface 41b of the main body portion 41. The insert 40 has a total of two support portions 42, equally spaced at intervals of 180° in the circumferential direction of the main body portion 41. Each support portion 42 extends radially outward from the main body portion 41.

[0065] The support portion 42 has an arm portion 421 extending radially outward from the main body portion 41, and an arc portion 422 formed in an arc shape at the tip of the arm portion 421. When the insert 40 is inserted in a fixed position in the well 33, the arc portion 422 straddles the corner of the well 33 and contacts the protruding portion 35. This allows the support portion 42 to support the insert 40 in the well 33 without making the arm portion 421 excessively long.

[0066] When the insert 40 is inserted into a fixed position in the well 33, the gaps formed between the main body 41 and the four corners of the well 33 are sampling portions 335. Two of the four sampling portions 335 are overlapped by the support portions 42.

[0067] 7, the distance between outer surface 41b and inner wall 33c of well 33 is relatively large in sampling section 335 compared to the location indicated by reference symbol 335x. Therefore, in sampling section 335, it is easier to insert an instrument such as a pipette compared to the location indicated by reference symbol 335x, and the contents in well 33 can be easily manipulated.

[0068] The insert 40 also has a protrusion 425 extending radially from the outer surface 41b of the main body portion 41. The protrusion 425 is provided at a position overlapping with the arm portion 421 of the support portion 42 in plan view.

[0069] 8 is a schematic cross-sectional view of the cell culture kit 200, and is a cross-sectional view taken along line VIII-VIII in FIG. 7. As shown in FIG. 8, the protrusion 425 extends to the vicinity of the inner wall 33c at the corner of the well 33 when the insert 40 is inserted in a fixed position in the well 33. Therefore, when the insert 40 tries to rotate in the circumferential direction of the insert 40 in the well 33, the protrusion 425 interferes with the inner wall 33c, and the rotation of the insert 40 in the circumferential direction is restricted. In this way, in the cell culture kit 200, the position of the insert 40 inserted in the well 33 can be stabilized.

[0070] Furthermore, the distance between the main body 41 and the inner wall 33c is kept constant by the two protrusions 425. This makes it possible to keep the position of the insert 40 constant within the well 33, making it easier to place the insert 40 at a fixed position within the well 33.

[0071] The cell culture kit 200 having such a configuration also becomes a novel cell culture kit in which the contents of the wells 33 can be easily manipulated.

[0072] Although the preferred embodiment of the present invention has been described above with reference to the accompanying drawings, the present invention is not limited to the above embodiment. The shapes and combinations of the components shown in the above embodiment are merely examples, and various modifications can be made based on the design, specifications, etc., without departing from the spirit of the present invention. [Explanation of symbols]

[0073] 10,30...well plate, 12b,122a...upper end, 13,33...well, 13c,33c...inner wall, 20,40...insert (cell culture insert), 21,41...main body, 21a...bottom, 21b,41b...outer surface, 22,42...support, 100,200...cell culture kit, 131...rectangle, 135,335...sampling part, 425...projection, VC...virtual circle, W1,W2...distance

Claims

1. A well plate having a plurality of wells; A cell culture insert is inserted into the well, The cell culture insert includes a cylindrical body having a bottom, a support portion extending radially outward from the main body portion at an upper portion of the main body portion, the support portion is provided at a position exposing at least a portion of the well when the cell culture insert is inserted into a fixed position in the well; A cell culture kit in which the well has a sampling portion in which, in the above-mentioned position, the distance between the outer surface of the main body portion and the inner wall of the well is relatively wide in the circumferential direction of the well.

2. The plurality of wells are arranged in a matrix in a plan view, The cell culture kit according to claim 1 , wherein the sampling portions of the plurality of wells face in the same direction in a plan view.

3. The body portion is cylindrical, The well has a corner protruding outward from an imaginary circle inscribed in the well, The cell culture kit according to claim 1 or 2, wherein the sampling portion is a gap formed between the main body portion and a corner of the well in the attitude.

4. The cell culture kit according to claim 3 , wherein the well has a rectangular shape in a plan view.

5. The cell culture insert has a plurality of the support portions provided discretely in a circumferential direction of the main body portion, 3. The cell culture kit according to claim 1, wherein an upper end of the well has a rotation-preventing portion that holds the support portion and determines the circumferential orientation of the cell culture insert.

6. The cell culture insert has a protrusion extending radially from an outer surface of the body portion, The cell culture kit according to claim 1 or 2, wherein the protrusion interferes with an inner wall of the well in the position.

Images