Holder for holding support used for cell culture

Abstract

To provide a holder with which cells can be observed in real time under a microscope and with which a support for cells after culturing can be readily collected from a culture medium and transferred.SOLUTION: A holder for holding a support used for cell culture, wherein the holder includes a plurality of recesses recessed from a planar surface toward a bottom surface in a plan view, a groove portion recessed from the planar surface toward the bottom surface in the plan view and provided so as to communicate with a part of each of the recesses, and one or more through-holes penetrating between the planar surface and the bottom surface, and each of the recesses has a side wall and includes an edge portion that extends in a direction away from the side wall on the bottom-surface side of the side wall and on which the support can be placed.SELECTED DRAWING: Figure 10

Description

[Technical Field]

[0001] The present invention relates to a holder for holding a support used for cell culture. [Background technology]

[0002] Plastic dishes are commonly used for cell culture. When observing the morphology of cells attached to these dishes, some microscopes can be difficult to focus and take clear morphological photographs.

[0003] To address this issue, glass-bottom dishes with a cover glass attached to the bottom of the dish are commercially available. While these dishes are suitable for real-time observation of cells under a microscope, the cover glass cannot be removed, making them unsuitable for culturing or immunostaining under consistent operating and culture conditions.

[0004] Patent Document 1 discloses a culture vessel with protrusions to make it easier to remove the cover glass with tweezers. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Patent Publication No. 9-117277 Summary of the Invention [Problem to be solved by the invention]

[0006] However, the culture vessel of Patent Document 1 has a problem in that the protrusions interfere with microscopic observation of the cells. [Means for solving the problem]

[0007] Therefore, the present inventors have developed a holder that allows cells to be observed under a microscope in real time and that also allows the cell support to be easily recovered and transferred from the culture medium after culture.

[0008] [1] An exemplary embodiment of the present invention is A holder for holding a support for cell culture, The above holder is a plurality of recesses recessed from the flat surface toward the bottom surface in a plan view; a groove portion provided so as to communicate with a part of each of the recesses and recessed from the plane surface toward the bottom surface in a plan view; one or more through holes penetrating between the flat surface and the bottom surface; Each of the recesses has a sidewall and an edge portion on the bottom side of the sidewall that extends in a direction away from the sidewall and on which the support can be placed. It is a holder.

[0009] By using the holder according to the above embodiment, cells can be observed under a microscope in real time, and the support for cells after culture can be easily recovered from the culture medium and transferred.

[0010] [2] In the holder according to [1], the plurality of recesses may be connected to one another via the grooves.

[0011] [3] [2] In the holder according to The plurality of recesses may be composed of one first recess and one or more second recesses, Each of the second recesses may be connected to the first recesses via the grooves.

[0012] [4] In the holder according to [3], each of the second recesses may have a bottom.

[0013] [5] In the holder according to [4], the first recess may be open and may be located at the center of the plurality of second recesses.

[0014] [6] In the holder according to [3], each of the second recesses may be open.

[0015] [7] The holder according to [1] may further include at least three legs that allow the height of the holder to be adjusted in the vertical direction.

[0016] [8] In the holder according to any one of [1] to [6], the through hole may be provided in the groove. [Brief explanation of the drawings]

[0017] [Figure 1] FIG. 1 shows a plan view of a holder 1 according to the first embodiment. [Figure 2] FIG. 2 shows a side view of the holder 1 according to the first embodiment. [Figure 3] FIG. 3 shows a cross-sectional view of the holder 1 taken along line AA in FIG. [Figure 4] FIG. 4 shows a perspective plan view of the holder 1 according to the first embodiment. [Figure 5] FIG. 5 shows a plan view of the holder 1 with the support 100 . [Figure 6] FIG. 6 shows a cross-sectional view of the holder 1 taken along line BB in FIG. [Figure 7] FIG. 7 shows an exploded view of the holder 1 with the support 100. [Figure 8] FIG. 8 shows a top perspective view of the holder 1 with three legs. [Figure 9] Figure 9(a) shows a side view of a holder 1 having three legs 60 submerged in a petri dish 300 containing culture medium 200, and Figure 9(b) shows a side view of the holder 1 with its height adjusted compared to the holder 1 in Figure 9(a). [Figure 10] FIG. 10 shows a perspective plan view of a holder 1A according to the second embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0018] definition For convenience, certain terms used in this application are collected here. Unless otherwise defined, all technical and scientific terms used in this application have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0019] The numerical ranges and parameters set forth herein are approximations; however, the numerical values ​​set forth in the specific examples are reported as precisely as possible. However, any numerical value inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Also, as used herein, the term "about" generally means within 10%, 5%, 1%, or 0.5% of a given value or range. Alternatively, the term "about" means within an acceptable standard error, as considered by one of ordinary skill in the art.

[0020] Hereinafter, embodiments of the present invention will be described. The following embodiments are merely examples, and the scope of the present invention is not limited to those shown in the following embodiments. Note that, to avoid repetition, explanations of similar content will be omitted as appropriate.

[0021] First embodiment FIG. 1 shows a plan view of a holder 1 according to this embodiment. FIG. 2 shows a side view of the holder 1 according to this embodiment. FIG. 3 shows a cross-sectional view of the holder 1 taken along line AA in FIG. 1. FIG. 4 shows a perspective plan view of the holder 1 according to this embodiment. FIG. 5 shows a plan view of the holder 1 including a support 100. FIG. 6 shows a cross-sectional view of the holder 1 taken along line BB in FIG. 5. FIG. 8 shows a perspective plan view of the holder 1 including three legs. FIG. 7 shows an exploded view of the holder 1 including the support 100. FIG. 8 shows a perspective plan view of the holder 1 including three legs. FIG. 9(a) shows a side view of the holder 1 including three legs 70 submerged in a Petri dish 300 containing a culture medium 200, and FIG. 9(b) shows a side view of the holder 1 with its height adjusted compared to the holder 1 in FIG. 9(a).

[0022] Holder 1 The holder 1 according to this embodiment is a device used in a cell culture vessel (e.g., a petri dish) and is capable of co-culturing two types of cells in one type of culture medium. The cell culture vessel is, for example, transparent. The holder 1 can detachably hold multiple supports 100. The supports 100 can culture and adhere cells on their surfaces. The support 100 is, for example, a thin glass member, which can also be called a cover glass. The holder 1 can have any shape (e.g., circular, polygonal, or elliptical) as long as it can be accommodated in the cell culture vessel, and is preferably circular in plan view. The holder 1 may have any thickness (e.g., 1 to 50 mm (millimeters)). The holder 1 has a plane 1a visible from the top side, a bottom surface 1b visible from the bottom side, and a side surface 1c visible from the side. The holder 1 may be made of a material such as a resin such as plastic (for example, polystyrene or polypropylene), glass or metal (for example, stainless steel), or may be made of any of these materials with a further surface treatment.

[0023] The holder 1 has a plurality of recesses 10 recessed from the flat surface 1a toward the bottom surface 1b in a plan view, and one or more through-holes 40 penetrating between the flat surface 1a and the bottom surface 1b. The recesses 10 can hold the support 100. By providing the holder 1 with the through-holes 40, for example, when the holder 1 is submerged in a cell culture vessel such as a petri dish containing a culture medium, substances secreted by cells into the culture medium can move between the flat surface 1a and the bottom surface 1b of the holder 1 through the through-holes 40.

[0024] Recess 10 The recesses 10 have a shape capable of accommodating the support 100. Preferably, each recess 10 has the same shape. The recesses 10 may be circular, polygonal, or elliptical in plan view. In one embodiment, the holder 1 includes one first recess 10a and multiple second recesses 10b. In one embodiment, each of the multiple second recesses 10b is arranged at equal intervals on a circumference centered on the first recess 10a. FIGS. 1 to 4 exemplarily show a holder 1 including one first recess 10a and six second recesses 10b (10b1, 10b2, 10b3, 10b4, 10b5, and 10b6).

[0025] In the holder 1 according to this embodiment, each of the recesses 10 has an opening 20, but each of the recesses 10 may have a bottom 80 (see FIG. 10 described later) (i.e., it does not have to be open). In one embodiment, each of the second recesses has a bottom 80, and the first recess is open (i.e., forms an opening 20). In another embodiment, each of the first recess and the second recess is open (i.e., has an opening 20). When the recesses 10 have openings 20, it becomes possible to observe the cells on the support 100 with a microscope while the holder 1 is submerged in a cell culture vessel.

[0026] Each recess 10 has a sidewall 50 extending from the flat surface 1a toward the bottom surface 1b. Each recess 10 has an edge 30 that extends away from the sidewall 50 (e.g., vertically) on the bottom surface 1b side of the sidewall 50 and on which the support 100 can be placed. In one embodiment, each recess 10 has an opening 20. In one embodiment, the edge 30 defines the opening 20. The size of the opening 20 can be changed by changing the length of the edge 30 from the sidewall 50. The flat surface 1a side has an upper surface 30a, and the upper surface 30a contacts the support 100 to hold it. That is, the support 100 can be placed on the edge 30 provided on the sidewall 50 as described above.

[0027] Groove 60 In one embodiment, the holder 1 has one or more grooves 60 recessed from the top surface 1a toward the bottom surface 1b in a plan view, and the multiple recesses 10 are connected to one another via the grooves 60. In one embodiment, each of the second recesses 10b is connected to the first recess 10a via the grooves 60. In one embodiment, the through-holes 40 are provided in the grooves 60 (i.e., in this embodiment, the grooves 60 also function as the through-holes 40). FIGS. 1 to 4 show six grooves 60, each of which has a through-hole 40, and each of the through-holes 40 is continuous with the openings 20 of the first recess 10a and the second recess 10b. Since the openings 20 are continuous with the through-holes 40, it is easy to retrieve the support 100 provided in the recess 10 using a tool (e.g., tweezers).

[0028] legs 60 In one embodiment, the holder 1 further includes at least three legs 70 that allow the height of the holder 1 to be adjusted vertically. By including at least three legs 70 in the holder 1, the height of the holder 1 from the bottom of the petri dish 300 containing the culture medium 200 can be stably and freely adjusted. In one embodiment, the legs 70 are screws. The holder 1 shown in FIGS. 9(a) and (b) includes three screw-type legs 70, and by rotating each of the screw-type legs 70, the height of the holder 1 from the petri dish 300 can be adjusted as shown in FIGS. 9(a) and (b).

[0029] Second embodiment 10 shows a perspective plan view of a holder 1A according to the second embodiment. The holder 1A according to this embodiment also allows cells to be observed under a microscope in real time, and allows the support 100 for cells after culture to be easily recovered and moved from the culture medium. That is, the holder 1A is a member capable of holding the support 100 used for cell culture, and has the following configuration. A plurality of recesses 10Ab (10Ab1 to 10Ab6) recessed from the flat surface toward the bottom surface in a plan view; A groove portion 60A is provided so as to communicate with a part of each of the recesses and is recessed from the plane surface toward the bottom surface in a plan view. One or more through holes 40A penetrating between the plane surface and the bottom surface; Each recess 10A has a side wall 50A, and an edge 30A on the bottom side of the side wall 50A that extends in a direction away from the side wall 50A and on which the support 100 can be placed.

[0030] Each of the recess 10Ab and the groove 60A may have a bottom surface or may be open. The holder 1A may further have a recess in the center as in the first embodiment.

[0031] Each through-hole 40A illustrated in Fig. 10 has a slit shape that exists on a line segment extending radially from the center of the holder 1A and between each recess 10Ab. With each through-hole 40A having such a radial slit shape, when the holder 1A is submerged in a cell culture vessel such as a petri dish containing culture medium, it is possible to suppress the occurrence of spontaneous stagnation of the culture medium and the occurrence of a concentration gradient when substances secreted into the culture medium by the cells move through the through-hole 40A between the flat surface 1Aa and the bottom surface (not shown) of the holder 1A. However, the shape of the through-hole 40A is not limited to the slit shape shown in Fig. 10 and may be, for example, a configuration in which multiple holes are arranged radially.

[0032] The position of the groove portion 60A that is provided to communicate with the recess 10Ab is not limited to the position closest to one end of the slit-shaped through hole 40A as illustrated in Figure 10, and it may be provided at another position as long as it can communicate with the recess 10Ab. [Explanation of symbols]

[0033] 1. 1A holder 1a, 1Aa plane 1b Bottom 1c, 1Ac side 10 recess 10a, 10Aa First recess 10b 1~6 , 10Ab 1~6 Second recess 20, 20A aperture 30, 30A edge 30a, 30aA top surface 40, 40A through hole 50, 50A side wall 60, 60A Groove 70 legs 100 support 200 Culture solution 300 Petri dishes

Claims

1. A holder for holding a support for cell culture, The holder is a plurality of recesses recessed from the flat surface toward the bottom surface in a plan view; a groove portion provided so as to communicate with a part of each of the recesses and recessed from the plane surface toward the bottom surface in a plan view; one or more through holes penetrating between the flat surface and the bottom surface; Each of the recesses has a sidewall and an edge portion on the bottom side of the sidewall that extends in a direction away from the sidewall and on which the support can be placed. holder.

2. The plurality of recesses are connected to each other via the grooves. The holder according to claim 1 .

3. the plurality of recesses are composed of one first recess and one or more second recesses, Each of the second recesses is connected to the first recesses via the grooves. The holder according to claim 2 .

4. Each of the second recesses has a bottom. The holder according to claim 3.

5. The first recess is open and is disposed at the center of the plurality of second recesses. The holder according to claim 4.

6. Each of the second recesses is open. The holder according to claim 3.

7. and at least three legs for vertically adjusting the height of the holder. The holder according to claim 1 .

8. The through hole is provided in the groove portion. A holder according to any one of claims 1 to 6.

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