Inserts and culture vessels with inserts
The insert with guide tube portions and notches in culture vessels addresses liquid overflow issues, ensuring efficient co-culture by managing capillary rise and enabling direct injection, with enhanced biocompatibility.
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- SCREEN HOLDINGS CO LTD
- Filing Date
- 2024-12-24
- Publication Date
- 2026-07-06
AI Technical Summary
The narrow spacing between wells in culture vessels and inserts leads to liquid rise due to capillary force, causing overflow and spreading, which is exacerbated by the use of multi-pipettes.
The insert features guide tube portions with notches and flange connections, along with a biocompatible gel, to manage liquid flow and prevent overflow, allowing direct access for injection devices.
Prevents liquid overflow and facilitates efficient co-culture by suppressing capillary rise and enabling direct injection, enhancing biocompatibility and biomimetic properties.
Smart Images
Figure 2026112123000001_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to an insert inserted into a well of a culture vessel for co-culture and a culture vessel with an insert having the insert.
Background Art
[0002] In recent years, for culturing cells mimicking organ functions, co-culture in which two or more types of cells are simultaneously cultured in one medium has been frequently performed. The cells cultured by co-culture are used, for example, in basic research on pharmaceuticals and toxicity evaluation. As a container for performing co-culture, a culture vessel having a well for holding a medium and an insert that is inserted into the well and arranged so as not to contact the bottom surface of the well are widely commercially available and used. Such an insert is disclosed in, for example, Patent Document 1. Patent Document 1 discloses a method for preparing a co-culture model in which the bottom surface of a 12-well culture plate is coated with a culture solution, cells are seeded and cultured therein, and another cell is seeded inside the insert (paragraph 0037 et al.).
Prior Art Documents
Patent Documents
[0003]
Patent Document 1
Summary of the Invention
Problems to be Solved by the Invention
[0004] As described in Patent Document 1, multi-well plates having multiple wells, such as 4, 8, 12, 24, or 48, are commonly used as culture vessels. Recently, multi-pipettes that can simultaneously inject culture medium (culture solution) or seed cells into multiple wells have become widely used. When using a multi-pipette, each well needs to be formed in the culture vessel at a spacing that matches each syringe of the multi-pipette. In this case, the spacing between adjacent wells becomes narrow, and the gap between the inner wall of each well and the insert inserted into the well also tends to become narrow. As a result, there is a risk that the liquid in the well may rise through the narrow gap between the inner wall of the well and the insert due to capillary force, rise to the top surface of the insert and spread, or overflow from the well.
[0005] This invention has been made in view of these circumstances, and aims to provide a technology that can suppress the rise of liquid in a well through the narrow gap between the inner wall of the well and the insert due to capillary force. [Means for solving the problem]
[0006] To solve the above problems, the first invention of the present application is an insert for insertion into a well of a culture vessel for co-culture, having a plurality of guide tube portions. Each of the plurality of guide tube portions has a cylindrical shape and is inserted downward along its central axis inside the well, and a guide opening is formed at its upper end for guiding an injection device for injecting liquid into the well. The gap between the outer surface of the guide tube portion and the inner wall of the well into which the guide tube portion is inserted is non-uniform when viewed in the circumferential direction about the central axis. The guide tube portion has a notch in a portion of the upper end of the outer surface, including a location where the gap is smallest.
[0007] The second invention of this application is an insert of the first invention, having a connecting portion which has flange portions that extend radially outward from the upper end of each of the multiple guide cylinder portions, and the flange portions connect to each other, thereby connecting the multiple guide cylinder portions. The connecting portion has a second notch located above the notch.
[0008] The third invention of this application is an insert according to the first or second invention, wherein the shape of the inner wall of the well in plan view is rectangular. Furthermore, the shape of the outer surface of the guide cylinder in plan view is circular. The notch is formed in a portion of the outer surface of the guide cylinder, including the portion closest to the long side of the inner wall of the well.
[0009] The fourth invention of this application is an insert of the third invention, wherein the notch is formed in a portion of the outer surface of the guide cylinder portion that includes the portion closest to one long side of the inner wall of the well, and in a portion that includes the portion closest to the other long side of the inner wall of the well.
[0010] The fifth invention of this application is an insert of the first invention, further comprising a connecting portion and an access hole. The connecting portion has a flange portion that extends radially outward from the upper end of each of the plurality of guide cylinder portions in a U-shape in plan view, and the plurality of guide cylinder portions are connected to each other by the flange portions connecting to each other. The access hole is formed inside the U-shape of the flange portion. When the plurality of guide cylinder portions are each inserted into one of the wells, the injection device can directly access the inside of the wells through the access hole.
[0011] The sixth invention of this application is an insert according to any one of the first to fifth inventions, further comprising a gel disposed at the opening at the lower end of each of the plurality of guide cylinder portions.
[0012] The seventh invention of this application is an insert of the sixth invention, wherein the gel is a cell-containing gel.
[0013] The eighth invention of this application is an insert according to the sixth or seventh invention, wherein the gel is formed by the curing of a thermosetting gel, and the solid component of the thermosetting gel is a biologically derived substance.
[0014] The ninth invention of this application is an insert according to any one of the sixth to eighth inventions, wherein the gel is formed by the curing of a thermosetting gel, and the solid component of the thermosetting gel is collagen.
[0015] The tenth invention of this application is an insert according to any one of the sixth to eighth inventions, wherein the gel is formed by the curing of a thermosetting gel, and the solid component of the thermosetting gel is fibrin.
[0016] The eleventh invention of this application is a culture vessel with an insert, comprising an insert according to any one of the first to tenth inventions, and the culture vessel having a plurality of bottomed cylindrical wells. [Effects of the Invention]
[0017] According to the first to eleventh inventions of this application, it is possible to suppress the rise of the liquid in the well to the upper end of the guide cylinder portion by capillary force through the gap between the inner wall of the well and the outer surface of the guide cylinder portion of the insert. Furthermore, it is possible to directly connect the liquid in the well and the liquid in the insert through the notch.
[0018] In particular, according to the second invention of this application, it is possible to further suppress the rise of the liquid in the well to the upper surface of the insert due to capillary force.
[0019] In particular, according to the fifth invention of this application, the injection device can be easily accessed to the liquid in the well through the U-shaped access hole on the inside of the insert. [Brief explanation of the drawing]
[0020] [Figure 1] It is a perspective view of a culture vessel with an insert. [Figure 2] It is a perspective view of a culture vessel. [Figure 3] It is a plan view of a culture vessel. [Figure 4] It is a perspective sectional view of a culture vessel with an insert. [Figure 5] It is a longitudinal sectional view of a culture vessel with an insert. [Figure 6] It is a perspective view of an insert. [Figure 7] It is a perspective view of an insert. [Figure 8] It is a schematic diagram showing a co-culture model that can be produced using a culture vessel with an insert. [Figure 9] It is a plan view of an insert placed on a culture vessel.Figure 1 is a perspective view of a culture vessel 1 with an insert according to one embodiment of the present invention. This culture vessel 1 with an insert is a device for culturing cells in a liquid L (culture medium) injected into each of a plurality of wells W provided in a culture vessel 10. In particular, the culture vessel 1 with an insert according to this embodiment allows for co-culturing of two or more types of cells simultaneously in each well W by inserting an insert 20 into each well W of the culture vessel 10 and culturing cells in the insert 20. As shown in Figure 1, the culture vessel 1 with an insert comprises a culture vessel 10 and an insert 20. Note that in Figure 1, for ease of understanding, only one insert 20 inserted into a well W of the culture vessel 10 is shown.
[0023] Figure 2 is a perspective view of the culture vessel 10. In Figure 2, for ease of understanding, one insert 20 inserted into the well W of the culture vessel 10 is shown with a dashed line. Figure 3 is a plan view of the culture vessel 10. As shown in Figures 2 and 3, the culture vessel 10 of this embodiment has a roughly rectangular three-dimensional shape. Furthermore, the shape of the culture vessel 10 of this embodiment in plan view is a roughly rectangular shape consisting of a pair of first longitudinal sides 101a, 101b and a pair of second transverse sides 102a, 102b. Hereinafter, the direction in which the first longitudinal sides 101a, 101b extend in the plan view of the culture vessel 10 will be referred to as the "first direction D1," and the direction in which the second longitudinal sides 102a, 102b extend will be referred to as the "second direction D2." The culture vessel 10 is made of, for example, resin.
[0024] The culture vessel 10 has a plurality of wells W arranged in a matrix when viewed from above in the vertical direction. In the culture vessel 10 of this embodiment, four wells W are arranged in a row in a first direction D1, and two rows of the four wells W arranged in the first direction D1 are arranged in a second direction D2. That is, the culture vessel 10 of this embodiment has a total of eight wells W. Each well W is a bottomed cylindrical shape that opens upward. The well W is a storage section capable of holding liquid L (see Figure 8 described later).
[0025] Figure 4 is a perspective cross-sectional view of the culture vessel 1 with inserts. For ease of understanding, only one insert 20 inserted into four wells W arranged in the first direction D1 is shown in Figure 4. Figure 5 is a longitudinal cross-sectional view of the culture vessel 1 with inserts. As shown in Figures 4 and 5, each well W in this embodiment has a roughly rectangular three-dimensional shape that is elongated in the first direction D1. That is, the shape of the inner wall 12 of each well W in plan view is roughly rectangular. In other words, the cross-section of each well W (the cut surface along a plane perpendicular to the depth direction D3 (vertical direction) of the well W) is roughly rectangular. The bottom surface 11 of the well W is flat.
[0026] However, the shape of the well W is not limited to this. For example, the shape of the inner wall 12 of each well W in plan view may be a perfect circle, an ellipse, or a polygon including a square, etc. That is, each well W may have a cylindrical or polygonal prism shape. Also, the culture vessel 10 may be a single-well plate having only one well W.
[0027] Furthermore, the diameter and depth of each well W are approximately a few millimeters to several tens of millimeters. In this embodiment, the distance between the centers of the four wells W arranged in the first direction D1 of the culture vessel 10 is set to approximately 18 mm, for example, so as to match the spacing between the syringes of the multipipette (not shown) that are simultaneously inserted into the four wells W. This multipipette corresponds to the "injection device" of the present invention. In addition, a predetermined amount of liquid L (culture medium) is held in each well W as a culture medium that provides a growth environment for cells, etc. In this embodiment, a sufficient amount of liquid L, for example, about 2.4 milliliters, is held in each well W for co-culture.
[0028] In other words, in the culture vessel 10, multiple wells W are arranged close to each other in a limited space, with spacing that matches the spacing between each syringe of the multipipette. Each well W also has sufficient capacity to hold the amount of liquid L required for culture (for example, 2.4 milliliters). Furthermore, as described above, in this embodiment, in order to increase the capacity of each well W in the limited space of the culture vessel 10, each well W is made into a roughly rectangular parallelepiped shape. When liquid L is injected into each well W, the liquid level of liquid L is located near the top of the guide cylinder portion 21 of the insert 20, which will be described later.
[0029] Furthermore, positioning protrusions 13 are provided at both ends of the culture container 10 in the first direction D1. In this embodiment, for each row of wells W arranged in the first direction D1, positioning protrusions 13 are provided at both ends of the culture container 10 in the first direction D1. Each positioning protrusion 13 protrudes upward in a columnar shape at the upper end of the culture container 10. In addition, a support base 14 is provided near the positioning protrusions 13 of the culture container 10, forming a part of the upper surface of the culture container 10.
[0030] The insert 20 is a component that is detachably placed on a culture vessel 10 for co-culture and inserted into multiple wells W. Figure 6 is a perspective view of the insert 20. Figure 7 is a perspective view of the insert 20 from a different direction than that shown in Figure 6. As shown in Figures 6 and 7, the insert 20 has multiple guide tube portions 21 arranged in a matrix when viewed from above in the vertical direction, a gel 22, a connecting portion 23, and a positioning recess 24. The gel 22 is shown only in Figure 8, which will be described later, and is omitted in the other drawings.
[0031] The number of guide tube portions 21 in one insert 20 corresponds to the number of wells W in each row of wells W arranged in the first direction D1 of the culture vessel 10. That is, in this embodiment, one insert 20 has four guide tube portions 21. In addition, inserts 20 are provided in proportion to the number of rows of wells W in the culture vessel 10. That is, in this embodiment, two inserts 20 are provided. However, these two inserts 20 may be connected to each other in the second direction D2. Furthermore, each guide tube portion 21 is inserted into one well W in the depth direction D3 (vertical direction) of the well W.
[0032] Each guide tube portion 21 has a cylindrical shape. Each guide tube portion 21 extends cylindrically along the central axis 90, with the central axis 90 as its center. In this embodiment, each guide tube portion 21 has a cylindrical shape. That is, the shape of the outer surface 211 of each guide tube portion 21 in a plan view is approximately circular. The central axis 90 of each guide tube portion 21 approximately coincides with the depth direction D3 (vertical direction) of the well W. That is, each guide tube portion 21 is inserted downward into the well W along the central axis 90. Each guide tube portion 21 is inserted into one well W of the culture vessel 10 and holds cells in the well W. The outer diameter of each guide tube portion 21 is smaller than the width in the first direction D1 and the width in the second direction D2 of one well W.
[0033] As shown in Figure 5, each guide cylinder portion 21 has a large diameter portion 212, a converging portion 213, and a small diameter portion 214. The large diameter portion 212 extends cylindrically along the central axis 90 in a portion including the upper end of each guide cylinder portion 21. The converging portion 213 has a hollow frustoconical shape (funnel shape) that gradually converges downward from the lower end of the large diameter portion 212. The small diameter portion 214 extends cylindrically downward along the central axis 90 from the lower end of the converging portion 213. The small diameter portion 214 forms a portion including the lower end of the guide cylinder portion 21. The inner diameter and outer diameter of the small diameter portion 214 are smaller than the inner diameter and outer diameter of the large diameter portion 212, respectively. In addition, the inner space of the large diameter portion 212, the inner space of the converging portion 213, and the inner space of the small diameter portion 214 are in vertical communication with each other.
[0034] However, the shape of the guide tube portion 21 is not limited to this. For example, the guide tube portion 21 may have a polygonal cylindrical shape extending along the central axis 90. That is, each of the multiple guide tube portions 21 has a cylindrical shape and is inserted downward along its own central axis 90 into the inside of the well W.
[0035] Furthermore, as shown in Figures 6 and 7, the insert 20 has multiple flange portions 25. The flange portions 25 extend substantially horizontally outward in the radial direction perpendicular to the central axis 90 from the upper end of each guide cylinder portion 21. The flange portions 25 also extend substantially horizontally at positions that do not cover the opening 210 at the upper end of each guide cylinder portion 21. The flange portions 25 connect to each other to form a connecting portion 23. The connecting portion 23 is located at the upper end of the insert 20 and is a member that extends substantially horizontally. In addition, the connection of the flange portions 25 allows multiple (four in this embodiment) guide cylinder portions 21 of a single insert 20 to be connected to each other.
[0036] Furthermore, positioning recesses 24 are formed at both ends of the connecting portion 23 in the first direction D1. Each positioning recess 24 is recessed inward in the first direction D1 at both ends of the connecting portion 23. When placing the connecting portion 23 of the insert 20 on the culture vessel 10, the positioning protrusions 13 provided on the culture vessel 10 are fitted into the positioning recesses 24 of the insert 20. This allows the insert 20 to be positioned in the appropriate location relative to the culture vessel 10. In addition, both ends of the connecting portion 23 of the insert 20 can be placed on the support base 14 of the culture vessel 10. This allows the connecting portion 23 of the insert 20 to be hooked onto and supported by the culture vessel 10.
[0037] Furthermore, with the connecting portion 23 of the insert 20 placed on the culture vessel 10, each of the multiple (four in this embodiment) guide tube portions 21 is inserted into one well W of the culture vessel 10. Also, when each guide tube portion 21 is inserted into one well W, it is positioned approximately in the center of the second direction D2 in one well W. In addition, with the connecting portion 23 of the insert 20 placed on the culture vessel 10, the lower end of each guide tube portion 21 is positioned above the bottom surface 11 of the well W.
[0038] Furthermore, with the connecting portion 23 of the insert 20 placed on the culture vessel 10, each syringe of the multipipette can be simultaneously inserted into the opening 210 at the upper end of each guide tube portion 21. Then, liquid L can be injected from each syringe into each well W through the opening 210 and the inner space of each guide tube portion 21. In other words, in this embodiment, an opening 210 is formed at the upper end of each of the multiple guide tube portions 21 of the insert 20, serving as a guide port to guide each syringe of the injection device for injecting liquid L into the well W.
[0039] Furthermore, each guide tube portion 21 has notches 50a and 50b. In this embodiment, two notches 50a and 50b are formed in each guide tube portion 21. The effects of forming notches 50a and 50b in each guide tube portion 21 will be described in detail later.
[0040] Furthermore, the flange portion 25 of this embodiment includes a plurality (four in this embodiment) of U-shaped portions 251. The U-shaped portions 251 are the parts of the flange portion 25 that spread out in a U-shape when viewed from above. When the connecting portion 23 of the insert 20 is placed on the culture vessel 10, each U-shaped portion 251 is located above one well W in the culture vessel 10 and is adjacent to the guide cylinder portion 21 in the first direction D1. The inner space of each U-shaped portion 251 becomes an access hole 250 that guides each syringe of the multipipette for injecting liquid L into the well W. In other words, in this embodiment, the access hole 250 is formed inside the U-shape of the flange portion 25.
[0041] This allows each syringe of the multipipette to be inserted into the access hole 250 without passing through the inner space of each guide tube portion 21. Then, liquid L can be directly injected from each syringe into each well W through each access hole 250. In other words, in this embodiment, when multiple guide tube portions 21 are each inserted into the inside of one well W, the injection device can directly access the inside of the well W through the access hole 250. This makes it possible to easily access the liquid in the well W with the injection device through the U-shaped access hole 250 inside the insert 20. However, the U-shaped portion 251 and the access hole 250 are not necessarily required.
[0042] Figure 8 is a schematic diagram showing a co-culture model that can be fabricated using the culture vessel 1 with inserts. As shown in Figure 8, the gel 22 is a component placed in the inner space of the small-diameter portion 214 of each guide tube portion 21. The gel 22 is positioned to close the opening 220 at the lower end of each guide tube portion 21. That is, the gel 22 is placed in the opening 220 at the lower end of each of the multiple guide tube portions 21. The gel 22 can also be formed, for example, by injecting a thermosetting gel into the inner space of the small-diameter portion 214 of each guide tube portion 21 and allowing it to harden.
[0043] The solid component of the thermosetting gel is preferably a bio-derived substance. By using a bio-derived substance, a highly biocompatible gel 22 can be formed. Therefore, regardless of whether cells are cultured on the top, inside, or bottom surface of the gel 22, bioreproducibility can be enhanced. In this embodiment, the solid component of the thermosetting gel is, for example, collagen. However, the solid component of the thermosetting gel may be something other than collagen, such as fibrin or agarose.
[0044] Furthermore, the thermosetting gel may contain living cells. That is, gel 22 may be a cell-containing gel. When forming a cell-containing gel, for example, a cell suspension gel in which living cells are dispersed as a liquid thermosetting gel can be injected into the inner space of the small diameter portion 214 of the guide tube portion 21. This makes it possible to form a gel 22 in which living cells are dispersed and fixed. When curing the thermosetting gel by heating, maintaining the thermosetting gel at 40°C or lower, more preferably 38°C or lower, can reduce the death of living cells. In addition, the liquid thermosetting gel may contain biochemical substances such as proteins or drugs. This makes it possible to give gel 22 a specific function.
[0045] Furthermore, the lower surface of the gel 22 is formed flush with the lower end of each guide tube portion 21. That is, the lower surface of the gel 22 and the lower end of each guide tube portion 21 are smoothly continuous without any steps. As a result, even if air bubbles are generated in the liquid L when it is injected into the well W, it is possible to reduce the amount of air bubbles that remain between the lower surface of the gel 22 and the lower end of each guide tube portion 21. Therefore, the impact on the cultured cells can be reduced.
[0046] With the connecting portion 23 of the insert 20 placed on top of the culture vessel 10, the lower surface of the gel 22 is positioned above the bottom surface 11 of the well W. As shown in Figure 8, when culturing cells using this culture vessel 1 with insert, cells C1 are cultured on the upper surface of the gel 22, and cells C2 are cultured on the lower surface of the gel 22. In this embodiment, cells C3 are also cultured inside the gel 22. Liquid L is appropriately injected into the inner and outer spaces of the guide tube portion 21 inside each well W. In this way, co-culture between cells C1 and C2 via the gel 22 can be achieved.
[0047] Thus, in this embodiment, by using an insert 20 with a gel 22, a co-culture model consisting of multiple cell types can be created without using a porous membrane. Furthermore, by using a highly biocompatible gel 22, it becomes possible to create a cell model with a high degree of biomimetic properties.
[0048] <2. Detailed configuration of the cutouts> Next, we will explain the detailed configuration of notches 50a and 50b.
[0049] Figure 9 is a plan view of one insert 20 placed on top of the culture vessel 10. In Figure 9, the culture vessel 10 is shown with a dashed line for ease of understanding. As described above, the shape of the inner wall 12 of each well W in plan view is approximately rectangular. Hereafter, in the plan view of each well W, the pair of longitudinal sides of the inner wall 12 will be referred to as "long sides 121a, 121b," and the pair of transverse sides will be referred to as "short sides 122a, 122b." The shape of the outer surface 211 of each guide tube portion 21 in plan view is approximately circular. Furthermore, when inserted into one well W, each guide tube portion 21 is located approximately in the center of the second direction D2 in one well W. Therefore, the gap Dx between the outer surface 211 of the guide cylinder portion 21 and the inner wall 12 of the well W into which the guide cylinder portion 21 is inserted is non-uniform when viewed in the circumferential direction (direction along the arc) with respect to the central axis 90 of the guide cylinder portion 21.
[0050] Furthermore, in the culture vessel 10, multiple wells W are arranged close to each other in the first direction D1 at intervals matching the spacing between each syringe of the multipipette. In addition, in the culture vessel 10, in order to provide more rows of wells W in a limited space, multiple wells W are arranged close to each other in the second direction D2. As a result, the width of each well W in the first direction D1 and the second direction D2 is small. Consequently, in a plan view, the gap Dx is smallest at the proximity points P1 and P2 where each of the pair of long sides 121a and 121b of the inner wall 12 of the well W and the outer surface 211 of the guide cylinder 21 inserted into the well W are closest in the second direction D2.
[0051] Therefore, as shown in Figures 6 and 7, in this embodiment, notches 50a and 50b are provided in a portion of the guide cylinder portion 21, including the upper end of each of the proximity points P1 and P2. That is, the guide cylinder portion 21 in this embodiment has notches 50a and 50b in a portion of the upper end of the outer surface 211, including the proximity points P1 and P2 where the gap with the inner wall 12 of the well W is minimized. More specifically, notch 50a is formed in a portion of the outer surface 211 of the guide cylinder portion 21, including the proximity point P1, which is closest to the long side portion 121a of the inner wall 12 of the well W. Notch 50b is formed in a portion of the outer surface 211 of the guide cylinder portion 21, including the proximity point P2, which is closest to the long side portion 121b of the inner wall 12 of the well W. Furthermore, the notches 50a and 50b each extend vertically from the upper end of the guide cylinder portion 21 to near the center of the large diameter portion 212 in the vertical direction. The notches 50a and 50b are holes that connect the inner space and the outer space of the guide cylinder portion 21.
[0052] In this way, by providing the notches 50a and 50b, the gap Dx between the outer surface 211 and the inner wall 12 of the well W can be widened in a portion of the outer surface 211 of the guide cylinder 21, including the upper end of the outer surface 211. This prevents the liquid in the well W from rising to the upper end of the guide cylinder 21 by capillary force through the narrow gap Dx between the inner wall 12 of the well W and the outer surface 211 of the guide cylinder 21. As a result, it is possible to prevent the liquid in the well W from rising and overflowing from the well W. In addition, the liquid L in the well W and the liquid L in the inner space of the guide cylinder 21 can be directly connected via the notches 50a and 50b. Figure 10 is a schematic diagram showing, for comparison purposes, how the liquid in the well W rises to the upper end of the guide cylinder 21 by capillary force through the gap Dx between the inner wall 12 of the well W and the outer surface 211 of the guide cylinder 21 when notches 50a and 50b are not provided in the guide cylinder 21.
[0053] Furthermore, in this embodiment, second notches 60a and 60b are formed in the connecting portion 23 of the insert 20 at locations above the notches 50a and 50b provided in the guide cylinder portion 21 (hereinafter referred to as "upper locations P3 and P4"). More specifically, a second notch 60a is formed in the upper location P3 of the connecting portion 23 of the insert 20, which is located above the notch 50a of the guide cylinder portion 21. Also, a second notch 60b is formed in the upper location P4 of the connecting portion 23 of the insert 20, which is located above the notch 50b of the guide cylinder portion 21. By providing the second notches 60a and 60b, the notches 50a and 50b can be opened further upward. This further suppresses the rise of the liquid L in the well W to the upper surface of the insert 20 due to capillary force. As a result, the movement of the liquid L in the well W to the outside of the well W via the upper surface of the insert 20 can be further suppressed.
[0054] <3. Variant> Although one embodiment of the present invention has been described above, the present invention is not limited to the above-described embodiment.
[0055] <3-1. First variation> In the above embodiment, the shape of the inner wall 12 of each well W in plan view was approximately rectangular, and the shape of the outer surface 211 of each guide cylinder portion 21 in plan view was approximately circular. However, as shown in the first modified example in Figure 11, the shape of the inner wall 12 of each well W in plan view may be approximately square, and the shape of the outer surface 211 of each guide cylinder portion 21 in plan view may be approximately circular. Furthermore, each guide cylinder portion 21 may be inserted into one well W and positioned approximately in the center of the first direction D1 and second direction D2 within one well W. Even in this case, the gap Dx between the outer surface 211 of the guide cylinder portion 21 and the inner wall 12 of the well W into which the guide cylinder portion 21 is inserted will be non-uniform when viewed in the circumferential direction (direction along the arc) centered on the central axis 90 of the guide cylinder portion 21.
[0056] Furthermore, in this modified example, the gap Dx is smallest at the proximity points P11, P12, P13, and P14, where each of the four sides of the inner wall 12 of the well W is closest to the outer surface 211 of the guide cylinder portion 21 inserted into the well W in a plan view. Therefore, in this modified example, a notch can be provided in a portion of the guide cylinder portion 21, including the upper end, at each of the proximity points P11, P12, P13, and P14.
[0057] This allows the gap Dx between the outer surface 211 and the inner wall 12 of the well W to be widened in some areas, including the upper end of the outer surface 211 of the guide cylinder 21, at adjacent locations P11, P12, P13, and P14. This prevents the liquid in the well W from rising to the upper end of the guide cylinder 21 by capillary force through the narrow gap Dx between the inner wall 12 of the well W and the outer surface 211 of the guide cylinder 21. As a result, it is possible to prevent the liquid in the well W from rising and overflowing from the well W.
[0058] <3-2. Second variation> Furthermore, as shown in the second modified example in Figure 12, the shape of the inner wall 12 of each well W in plan view may be approximately circular, and the shape of the outer surface 211 of each guide cylinder portion 21 in plan view may be approximately square. Each guide cylinder portion 21 may be inserted into one well W and positioned approximately in the center of the first direction D1 and the second direction D2 within that well W. Even in this case, the gap Dx between the outer surface 211 of the guide cylinder portion 21 and the inner wall 12 of the well W into which the guide cylinder portion 21 is inserted will be non-uniform when viewed in the circumferential direction (direction along the arc) with respect to the central axis 90 of the guide cylinder portion 21.
[0059] Furthermore, in this modified example, the gap Dx is smallest at the proximity points P21, P22, P23, and P24 (the four corners of the outer surface 211 of the guide cylinder 21 in a plan view) where the inner wall 12 of the well W and each of the four sides of the outer surface 211 of the guide cylinder 21 inserted into the well W are in the closest proximity. Therefore, in this modified example, notches can be provided in a portion of the guide cylinder 21, including the upper end, at each of the proximity points P21, P22, P23, and P24.
[0060] This allows the gap Dx between the outer surface 211 and the inner wall 12 of the well W to be widened in some areas, including the upper end of the outer surface 211 of the guide cylinder 21, at adjacent locations P21, P22, P23, and P24. This prevents the liquid in the well W from rising to the upper end of the guide cylinder 21 by capillary force through the narrow gap Dx between the inner wall 12 of the well W and the outer surface 211 of the guide cylinder 21. As a result, it is possible to prevent the liquid in the well W from rising and overflowing from the well W.
[0061] Furthermore, the elements that appear in the above embodiments and modifications may be combined or omitted as desired, as long as no inconsistencies arise. [Explanation of Symbols]
[0062] 1: Culture vessel with insert 10:Culture container 11: Bottom (of the culture vessel) 12: Inner wall (of the culture vessel) 13: Positioning protrusion 14: Support stand 20: Insert 21: Guide tube section 22: Gel 23:Connection part 24: Positioning recess 25: Flange section 50a: Notch 50b: Notch 60a: Second notch 60b: Second notch 90: (The central axis of the guide tube) 121a: Long side (of a well) 121b: Long side (of a well) 122a: Short side (of the well) 122b: Short side (of a well) 210: Opening at the upper end (guide port) of the guide tube. 211: Outer surface (of the guide tube) 250: Access port 251: U-shaped section Dx: Gap L: Liquid used as a culture medium (culture solution) W: Well
Claims
1. An insert that is inserted into the wells of a culture vessel for co-culture, Each of the following guide tubes has a cylindrical shape, is inserted downward along its central axis inside the well, and has a guide opening formed at its upper end for guiding an injection device for injecting liquid into the well. It has, The gap between the outer surface of the guide cylinder and the inner wall of the well into which the guide cylinder is inserted is non-uniform when viewed in the circumferential direction with respect to the central axis. The guide tube portion is an insert having a notch in a part of the upper end of the outer surface, including the location where the gap is smallest.
2. The insert according to claim 1, A connecting part having flange portions that extend radially outward from the upper end of each of the multiple guide cylinder portions, and the flange portions connect to each other, thereby connecting the multiple guide cylinder portions to each other. It has, The connecting portion is an insert having a second notch located above the aforementioned notch.
3. An insert according to claim 1 or claim 2, The inner wall of the well has a rectangular shape in plan view. The outer surface of the guide cylinder portion, when viewed from above, has a circular shape. The notch is formed in a portion of the outer surface of the guide cylinder portion, including the portion closest to the long side of the inner wall of the well, in the insert.
4. The insert according to claim 3, The insert is formed in which the notches are located on the outer surface of the guide cylinder portion, specifically in a portion that includes the location closest to one of the long sides of the inner wall of the well, and in a portion that includes the location closest to the other long side of the inner wall of the well.
5. The insert according to claim 1, Each of the multiple guide cylinders has a flange portion that extends radially outward from its upper end, having a U-shape in plan view, and the flange portions connect to each other, thereby linking the multiple guide cylinders together; An access hole is formed on the inside of the U-shaped flange portion, It further possesses, When multiple guide tubes are each inserted into the inside of one of the wells, the injection device is an insert that allows direct access to the inside of the well through the access hole.
6. An insert according to claim 1 or claim 2, Gel placed at the opening at the lower end of each of the multiple guide tube sections An insert further possessing...
7. The insert according to claim 6, The aforementioned gel is a cell-containing gel, an insert.
8. The insert according to claim 6, The aforementioned gel is formed by the curing of a thermosetting gel, and the solid component of the thermosetting gel is a bio-derived substance, which is the insert.
9. The insert according to claim 6, The aforementioned gel is formed by the curing of a thermosetting gel, and the solid component of the thermosetting gel is collagen, in the insert.
10. The insert according to claim 6, The gel is formed by the curing of a thermosetting gel, and the solid component of the thermosetting gel is fibrin, in the insert.
11. The insert according to claim 1 or claim 2, The culture vessel having a plurality of bottomed cylindrical wells, A culture vessel with an insert, having the following features.