JP2016129495A - Cell culture vessel - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a cell culture vessel suitable for observing a cultured cell.SOLUTION: In a cell culture vessel 100, a housing space 140 which is open above and surrounded with a peripheral wall part 120 erected from a vessel bottom part 110 is formed, the peripheral wall part 120 has an inner wall surface 123 facing the housing space 140 and an outer wall surface 124 facing outside the housing space 140, and the inner wall surface 123 and the outer wall surface 124 cross at an opening side end.SELECTED DRAWING: Figure 1B

Description

  The present invention relates to a cell culture container suitable for observation of cultured cells.

  The in vitro fertilized egg (zygote) is produced by fertilizing sperm and ovum in a culture system, and the fertilized egg goes through the cleavage, morula, and blastocyst stages, and then emerges from the zona pellucida It is possible to culture up to the blastocyst stage. Assistive reproductive technology (ART) to transfer a fertilized egg from the cleavage to the blastocyst stage to the uterus to give birth to a baby is not limited to the livestock region. Established in infertility medicine.

  In in vitro fertilization, a microdrop method is often used in which a culture solution is dropped in a container, and a fertilized egg is placed in the container and cultured in vitro. Conventionally, in this microdrop method, a petri dish having a single flat bottom and a diameter of 30 to 60 mm is used as a cell culture container, and a plurality of drops of culture solution are produced on the bottom of the petri dish at intervals. A method has been used. However, when a drop is created with an ordinary petri dish, there is a problem that the drop formation position is not fixed and the drop is displaced due to vibration or the like. When the drop is shifted, there is a problem that it becomes difficult to identify a fertilized egg cultured and observed in the drop. Therefore, Patent Document 1 discloses a cell culture container that can control the position of the drop and suppress the influence of vibration during fertilized egg culture work or culture.

  Patent Document 1 discloses a cell culture container 800 shown in FIGS. 8A and 8B. The cell culture container 800 includes a container bottom portion 810, an outer peripheral wall portion 820 that stands up from the periphery of the container bottom portion 810, and a peripheral wall portion 830 that stands up from the container bottom portion 810. A space that is opened upward and is surrounded by the peripheral wall portion 830 is a storage space 840 for storing cells and a culture solution. On the surface forming the bottom of the accommodation space 840, a plurality of depressions 850 for positioning cells are formed. By using the cell culture container 800 and holding the microdrop of the culture solution in the accommodation space 840 and placing the cells in the depression 850, the positions of the microdrops and the cells can be fixed.

Japanese Patent No. 4724854

  The peripheral wall 830 in the conventional cell culture vessel 800 connects the inner wall surface 831 facing the accommodation space 840, the outer wall surface 832 facing the outside of the accommodation space 800, and the opening side ends of the inner wall surface 831 and the outer wall surface 832. And an upper surface 833 to be used.

  When culture is performed by holding a drop of a culture solution containing cells in the accommodation space 840 surrounded by the peripheral wall portion 830 of the cell culture container 800, a microscope is used to observe the inside of the accommodation space 800. However, if an object to be observed is searched from the entire surface of the cell culture vessel 800, the object may be held on the upper surface 833 of the peripheral wall portion 830. Therefore, the bottom of the accommodation space 800 where the cells are arranged is disposed. Therefore, it was necessary to observe the upper surface 833 by shifting the focus.

  The target object to be observed is not limited to cells or the like to be cultured, but may be a foreign object. Foreign body management is very important in cell culture. This is because cells may be affected in various ways by foreign substances present during cell culture, resulting in growth inhibition, loss of function, and changes in characteristics. The presence of foreign matter also affects the quality evaluation of cells. In particular, it interferes with optical observation with a microscope or the like. Furthermore, it becomes a data noise factor when analyzing the culture medium. Furthermore, when culturing cells for the purpose of regenerative medicine and infertility treatment, it is considered that foreign substances are transplanted together with cells, which is a big problem. When the size of the foreign matter is small, for example, the entire storage space 800 is observed with a microscope.

  Therefore, the present invention is a cell culture container in which a storage space for storing cells and culture medium surrounded by a peripheral wall portion is formed, and there is a possibility of unintentional retention of cells and foreign substances on the surface of the peripheral wall portion. It is an object of the present invention to provide a cell culture container with reduced slag.

Therefore, the present invention discloses the following invention as means for solving the above-described problems.
(1)
A cell culture container in which an accommodation space for accommodating cells and a culture solution is formed,
The housing space is a space that is surrounded by a peripheral wall portion that stands up from the bottom of the container and opens upward.
The peripheral wall portion has an inner wall surface facing the housing space and an outer wall surface facing the outer side of the housing space,
The cell culture container, wherein the inner wall surface and the outer wall surface intersect at an opening side end.
(2)
The cell culture container according to (1) above, wherein a portion where the inner wall surface and the outer wall surface intersect is a continuous line surrounding the opening of the accommodation space in a plan view.
According to the invention of (1) above, since there is no upper surface at the opening side end of the peripheral wall portion, there is a low possibility that cells and foreign substances are held on the surface of the peripheral wall portion.
According to the invention of (2) above, since the opening side end of the peripheral wall portion forms a linear top having substantially no width in plan view, cells and foreign substances are held at the opening end of the peripheral wall portion. Is less likely to be done.

  According to the present invention, a cell culture container in which a storage space for storing cells and culture medium surrounded by a peripheral wall portion is formed, and it is possible to unintentionally hold cells and foreign substances on the surface of the peripheral wall portion. A cell culture container with reduced properties is provided.

It is a schematic diagram of the top view of the cell culture container 100 which concerns on Embodiment 1 of this invention. It is a schematic diagram of the XX section of cell culture container 100 shown in Drawing 1A. It is a schematic diagram of the cross section of the cell culture container 100 which concerns on Embodiment 2 of this invention. It is a schematic diagram of the cross section of the cell culture container 100 which concerns on Embodiment 3 of this invention. It is a schematic diagram of the cross section of the cell culture container 100 which concerns on Embodiment 4 of this invention. It is a schematic diagram of the cross section of the cell culture container 100 which concerns on Embodiment 5 of this invention. It is a schematic diagram of the cross section of the cell culture container 600 which is a comparative example. It is a schematic diagram of the top view of the cell culture container 100 which concerns on Embodiment 6 of this invention. It is the schematic diagram of the XX cross section of the cell culture container 100 shown to FIG. 7A. It is a schematic diagram of the top view of the conventional cell culture container 800 currently disclosed by patent document 1. FIG. It is the schematic diagram of the YY cross section of the conventional cell culture container 800 shown to FIG. 8A. It is a figure explaining the definition of the inclination-angle of a surface. It is a schematic diagram of the end surface of the surrounding wall part 120 along the plane along an up-down direction. It is a figure for demonstrating the effect of Embodiment 3. FIG. It is a schematic diagram of the surrounding wall part 120 in the modification 1 of Embodiment 4. FIG. It is a schematic diagram of the surrounding wall part 120 in the modification 2 of Embodiment 4. FIG.

  The cell culture container of the present invention will be described with reference to the embodiments shown in the drawings, but the scope of the present invention is not limited to these embodiments. In the drawings of the present specification, configurations having the same functions are denoted by the same reference numerals regardless of the dimensions and shapes of the respective configurations, and description thereof is omitted except for particular differences.

<Inclination angle>
Before describing details of Embodiments 1 to 7 described later, the inclination angles between the inner wall surface and the outer wall surface of the peripheral wall portion will be described. The reference numerals referred to in the following description are the same as the reference numerals for explaining the first to seventh embodiments.

  In the present invention, the “vertical direction” means the depth direction of the liquid (that is, the vertical direction) when a liquid such as a culture solution is stored in the storage space 140 in a state where the cell culture vessel 100 of the present invention is placed on a horizontal plane. ). And in this invention, let the direction which goes to an opening from the bottom of the accommodation space 140 along an up-down direction be upper direction, and let the direction which goes to the bottom from the opening of an accommodation space along an up-down direction be the downward direction.

In the present invention, the inclination angle of the surface is defined as follows. As shown in FIG. 9, N is a normal line passing through a point P on the surface S that is the surface of the member X, and L is a virtual straight line along the vertical direction (as defined above) passing through the point P. A normal line N and an imaginary straight line when a half line part of the normal line N from the point P where the member X does not exist is N ₁ and a half line part above the point P of the virtual line L is L _1. Of the angles formed by L, the angle θ ₁ formed between the half-line portion N ₁ , the point P, and the half-line portion L ₁ is defined as the inclination angle at the position of the point P on the surface S. . The inclination angle of the whole or a part of the inner wall surface 123 or the outer wall surface 124 of the peripheral wall surface 120 is a point included in the whole or a part thereof, the point P is the peripheral wall portion 120, and the inner wall surface 123 or the outer wall surface. The inclination angle θ ₁ at the position of the point P defined above when the surface S is 124 is indicated.

  The cell culture container 100 of the present invention includes a peripheral wall 120. The peripheral wall 120 has an inner wall surface 123 that faces the housing space 140 and an outer wall surface 124 that faces the outside of the housing space 140.

  FIG. 10 shows a schematic diagram of the end surface of the peripheral wall 120 by a plane passing through the center of gravity of the accommodation space when viewed in plan with the cell culture container 100 of the present invention placed on a horizontal plane.

  The inclination angle of the inner wall surface 123 of the peripheral wall 120 is preferably greater than 0 °, more preferably 10 ° or more, particularly preferably 30 ° or more, and preferably 90 ° or less at the opening side end or in the vicinity thereof. More preferably, it is 75 ° or less, particularly preferably 65 ° or less. However, at the opening side end 150, the inclination angle of the inner wall surface 123 is preferably 0 °. When the inclination angle at the opening side end of the outer wall surface 124 or in the vicinity thereof is 90 °, the inclination angle at the opening side end of the inner wall surface 123 or in the vicinity thereof is preferably less than 90 °. The inner wall surface 123 of the peripheral wall portion 120 extends to the opening side end at a substantially constant inclination at or near the opening side end (the first, second, third, fifth, seventh, fourth modification example 2). Alternatively, it is preferable that the inclination becomes smaller as it approaches the opening side end (the fourth embodiment, the first modification of the fourth embodiment, etc.).

  The inclination angle of the outer wall surface 124 of the peripheral wall portion 120 is preferably greater than 0 °, more preferably 10 ° or more, particularly preferably 30 ° or more, and preferably 90 ° or less, at or near the opening side end. More preferably, it is 75 ° or less, particularly preferably 65 ° or less. However, at the opening side end 150, the inclination of the outer wall surface 124 is preferably 0 °. When the inclination angle at the opening side end of the inner wall surface 123 or in the vicinity thereof is 90 °, the inclination angle at the opening side end of the outer wall surface 124 or in the vicinity thereof is preferably less than 90 °. The outer wall surface 124 of the peripheral wall 120 extends to the opening side end with a substantially constant inclination at or near the opening side end (the first, second, third, fifth, seventh, fourth modification example 2). Alternatively, it is preferable that the inclination becomes smaller as it approaches the opening side end (the fourth embodiment, the first modification of the fourth embodiment, etc.).

  In the present invention, "the opening side end or the vicinity thereof" of the inner wall surface or the outer wall surface, for example, when the width along the vertical direction from the bottom side end to the opening side end of the inner wall surface or the outer wall surface is 100, It is preferably 20 or less, more preferably 10 or less, preferably 3 or more, more preferably 5 or more along the vertical direction from the opening side end to the bottom side end of the inner wall surface or outer wall surface.

  The inclination angle of the portion closer to the bottom end than the vicinity of the opening end of the inner wall surface 123 of the peripheral wall portion 120 is not particularly limited, and is preferably 0 ° or more, more preferably greater than 0 °, more preferably It is 10 ° or more, particularly preferably 30 ° or more, preferably 90 ° or less, more preferably less than 90 °, more preferably 75 ° or less, and particularly preferably 65 ° or less.

  The inclination angle of the portion closer to the bottom end than the vicinity of the opening end of the outer wall surface 124 of the peripheral wall 120 is not particularly limited, and is, for example, 0 ° or more, preferably greater than 0 °, more preferably 10 °. More preferably, it is 30 ° or more, preferably 140 ° or less, more preferably 120 ° or less, more preferably 90 ° or less, more preferably less than 90 °, more preferably Is 75 ° or less, particularly preferably 65 ° or less.

  Each of the inner wall surface 123 and the outer wall surface 124 may have a substantially constant inclination angle from the bottom end toward the opening end (for example, Embodiments 1, 2, 3, and 7), or from the bottom end to the opening side. The tilt angle may be changed continuously or stepwise toward the end (for example, Embodiments 4 and 5). Alternatively, one or a plurality of stepped portions rising in a step shape may be formed at portions other than the opening side end of the inner wall surface 123 and / or the outer wall surface 124 or the vicinity thereof (for example, the outer wall surface of the second embodiment) 124).

  Embodiments of the present invention will be described below with reference to the drawings.

<Embodiment 1 (FIGS. 1A and 1B)>
1A and 1B show Embodiment 1 of the cell culture container 100 of the present invention.
The cell culture container 100 includes a container bottom part 110 and an outer peripheral wall part 130 erected on the periphery of the container bottom part 110, and further includes a peripheral wall part 120 erected from the container bottom part 110.

  The accommodation space 140 is a space for accommodating cells and a culture solution, and is a space that is surrounded by the peripheral wall 120 and opened upward.

  In the first embodiment, a portion of the container bottom portion 110 that forms the bottom of the storage space 140 is referred to as a storage space bottom portion 111, and a portion that connects the peripheral wall portion 120 and the outer peripheral wall portion 130 is referred to as an outer edge bottom portion 112.

  In the first embodiment, the peripheral wall 120 is formed by combining an inverted conical inner wall 121 and a cylindrical outer wall 122 connected to the upper end of the inner wall and extending downward. It is comprised so that it may exist so that between may become a hollow structure. However, the hollow structure is not a necessary configuration, and the entire peripheral wall portion 120 may have a solid structure like the peripheral wall portion 120 shown in FIGS.

  The number of the accommodation spaces 140 included in one cell culture container 100 is only 1 in the illustrated example, but may be 2 or more, for example, 2 to 4.

  The shape of the figure when the inner peripheral side contour of the upper end that is the open end of the outer peripheral wall 130 is viewed in plan can be an arbitrary shape such as a circle (including a circle and an ellipse), preferably It is circular, and its opening width (the maximum value of the distance between a pair of points on the periphery of the figure facing the center of gravity of the figure in the figure) is preferably 30 to 60 mm.

Next, the features of the peripheral wall 120 will be described in detail.
In the cell culture container 100 of the present invention, the peripheral wall 120 has an inner wall surface 123 facing the accommodation space 140 and an outer wall surface 124 facing the outside of the accommodation space 140.

  Here, the inner wall surface facing the accommodation space is a surface surrounding the accommodation space among the side surfaces of the peripheral wall portion, and is located inside the accommodation space when viewed in plan with the cell culture container of the present invention placed on a horizontal plane. It is the facing side. The outer wall surface facing the outside of the housing space is the surface facing the outside of the housing space when viewed in plan with the cell culture container of the present invention placed on a horizontal surface, among the side surfaces of the peripheral wall portion. Both the inner wall surface and the outer wall surface are perpendicular to the virtual straight line with respect to the virtual straight line passing through the center of gravity of the accommodation space when viewed in plan with the cell culture container of the present invention placed on a horizontal plane. Is projected as a line rather than a point. In the illustrated embodiment, the inner wall surface 123 does not include a surface facing the outside of the accommodation space 140, and the outer wall surface 124 does not include a surface facing the accommodation space 140.

  The inner wall surface 123 and the outer wall surface 124 intersect at the opening side end. Here, that the inner wall surface 123 and the outer wall surface 124 “intersect” at the opening side end means that the center of gravity of the accommodation space when the cell culture vessel 100 of the present invention is viewed in a plane in a state of being placed on a horizontal plane. It means that the inner wall surface 123 and the outer wall surface 124 substantially intersect at the opening side end on the cross section of the peripheral wall portion 120 along the vertical direction. “Substantially intersecting” means that the inner wall surface 123 and the outer wall surface 124 intersect at the opening side end on the cross section, or are connected through a surface having substantially no width. “Substantially no width” is as described in the next paragraph. With this configuration, at the time of cell culture, cells, foreign matters, and the like cannot be stably held on the opening side end of the peripheral wall portion 120 and fall along either the inner wall surface 123 or the outer wall surface 124. For this reason, possibility that a cell, a foreign material, etc. will remain in the state on the opening side end of the surrounding wall part 120 at the time of cell culture is reduced.

More preferably, as shown in FIGS. 1A and 1B, the portion where the inner wall surface 123 and the outer wall surface 124 intersect each other is seen in a plan view with the cell culture vessel 100 of the present invention placed on a horizontal plane (in the present invention, In this case, the line 150 is a continuous line 150 that surrounds the opening of the accommodation space 140. The line 150 is formed except when the inclination angle at the opening side end of the inner wall surface 123 and the inclination angle at the opening side end of the outer wall surface 124 are both 0 °. The slope of the surface changes discontinuously at the line 150. At this time, the surface of the opening side end of the peripheral wall portion 120 is a cross-section by a virtual plane passing through the center of gravity of the accommodation space 140 when viewed in plan with the cell culture vessel 100 placed on a horizontal plane, The line 150 is bent at the apex. Line 150 has substantially no width. Here, “substantially has no width” means that there is no width or a sufficiently small width in a plan view as compared with the cell to be cultured, for example, 1/2 of the width of the cell to be cultured. Below, preferably 1/10 or less, more preferably 1/20. For example, in the cell culture container 100 for culturing egg cells or embryos, the line 150 has a width of 50 μm or less, preferably 10 μm or less in plan view. Also good. In this case, it is preferable because the possibility that cells, foreign substances, and the like are held on the opening side end of the peripheral wall 120 during cell culture is greatly reduced. In a further preferred embodiment, when the inner wall surface 123 and the outer wall surface 124 intersect to form a line 150, the cell culture vessel 100 passes through the center of gravity of the accommodation space 140 when viewed in plan with being placed on a horizontal plane. An angle θ ₂ (see FIG. 10) formed in the peripheral wall portion 120 formed by the inner wall surface 123 and the outer wall surface 124 at the line 150 on the cross section by the plane along the vertical direction is preferably 90 ° or less, and more The angle is preferably 45 ° or less, more preferably 20 ° or less, and particularly preferably 10 ° or less. The smaller the θ _{2, the} sharper the opening side end of the peripheral wall 120, and the possibility that cells, foreign substances, etc. are held on the opening end of the peripheral wall 120 during cell culture is further reduced.

  In addition, the intersection part of the inner wall surface 123 and the outer wall surface 124 may be appropriately chamfered. At this time, the intersection between the inner wall surface 123 and the outer wall surface 124 is preferably chamfered so that a flat surface perpendicular to the vertical direction is not formed, and is chamfered so that a rounded surface is formed. It is more preferable. The rounded surface of the intersecting portion when chamfered is a cross section by a virtual plane passing through the center of gravity of the accommodation space 140 when viewed in plan with the cell culture vessel 100 placed on a horizontal plane. , The curvature radius is preferably 100 μm or less, more preferably 50 μm or less, and particularly preferably a surface that can be approximated to a part of an arc of 10 μm or less. At this time, it is preferable because cells, foreign substances, and the like are unlikely to be held at the intersections between the inner wall surface 123 and the outer wall surface 124.

  In Embodiment 1, the inclination angle at the opening side end of the inner wall surface 123 of the peripheral wall portion 120 and the vicinity thereof, and the inclination at the portion closer to the bottom end than the vicinity of the opening side end of the inner wall surface 123 of the peripheral wall portion 120 A preferable range of the angle is as described in <Inclination angle>. More preferably, the inclination angle of the inner wall surface 123 is within the range given as a preferable range of the inclination angle at the opening side end of the inner wall surface 123 and in the vicinity thereof from the bottom end to the opening end. In the illustrated embodiment 1, the inner wall surface 123 is formed so that the accommodation space 140 extends from the bottom toward the opening, and the inclination angle of the inner wall surface 123 is substantially constant from the bottom end to the opening end. However, it is not limited to this form.

  The inner wall surface 123 preferably includes a conical or frustoconical portion. The conical or frustoconical portion is formed on the bottom side of the culture vessel so that the apex of the cone or the upper surface and the lower surface of the truncated cone has the smaller area. Conical shapes include cones and elliptical cones, and similar shapes, such as cones or elliptical cones with rounded vertices, conical surfaces bulging outwards, conical surfaces recessed inwards Shape etc. are included. The truncated cone has a truncated cone and an elliptical frustum, and similar shapes, for example, a shape in which the junction between the upper or lower surface of the truncated cone or the truncated truncated cone and the truncated cone is rounded, and the truncated cone is on the outer side. And a shape having a conical surface recessed inward. The inner wall surface 123 is not limited to the above, and may have a polygonal pyramid shape, a polygonal frustum shape, or the like.

  In the first embodiment, the inclination angle at the opening side end of the outer wall surface 124 of the peripheral wall portion 120 and the vicinity thereof, and the inclination at the portion closer to the bottom end than the vicinity of the opening side end of the outer wall surface 124 of the peripheral wall portion 120. The preferable range of the angle is as described in <Inclination angle>. More preferably, the entire inclination angle of the outer wall surface 124 from the bottom side end to the opening side end is within the range mentioned as the preferable range of the inclination angle at the opening side end of the outer wall surface 124 and in the vicinity thereof. In the illustrated embodiment 1, the inclination angle of the outer wall surface 124 is substantially constant from the bottom end to the opening end, but is not limited to this configuration.

  When the inclination angles of the inner wall surface 123 and the outer wall surface 124 of the peripheral wall portion 120 are in the above range, the possibility that cells, foreign substances, etc. are held on the opening side end of the peripheral wall portion 120 during cell culture is particularly high. It can be effectively reduced.

  In the first embodiment, the magnitude relationship between the inclination angles of the inner wall surface 123 and the outer wall surface 124 is not particularly limited. As a typical example, the inclination angle of the inner wall surface 123 is at least at the opening side end and in the vicinity thereof. In this case, the inclination angle at the opening side end of the inner wall surface 123 and in the vicinity thereof is preferably 10 ° or more, more preferably 20 ° or more, particularly preferably 30 ° or more, preferably Is 70 ° or less, more preferably 60 ° or less, particularly preferably 50 ° or less, and the inclination angle at the opening side end of the outer wall surface 124 and its vicinity is preferably 30 ° or more, more preferably 50 ° or more, preferably Is 90 ° or less, more preferably 85 ° or less, and particularly preferably 80 ° or less.

  The bottom of the accommodation space 140 is formed by the accommodation space bottom surface 113 which is the upper surface of the accommodation space bottom 111. The cell culture container 100 is usually installed and used such that the accommodation space bottom surface 113 is horizontal. The shape of the periphery of the accommodation space bottom surface 113 and the opening side end of the peripheral wall 120 (same as the line 150 in the first embodiment) when viewed in plan along the vertical direction is not particularly limited, but is preferably similar to each other, preferably Are each circular (including circular and elliptical).

  In the present invention, the bottom surface of the accommodation space is preferably a flat surface, but is not limited thereto. When the bottom surface is not a plane, the plane including the peripheral line of the bottom surface is regarded as the “accommodating space bottom surface”. Examples of the non-planar bottom surface include a convex surface whose central portion protrudes upward and a concave surface whose central portion is depressed downward.

The accommodation space 140 is configured to accommodate a small volume of culture solution, for example, a drop of 10 μl to 100 μl of the culture solution, and the area of the accommodation space bottom surface 113 is accommodated by the drop of the culture solution of 10 μl to 100 μl. The entire surface of the bottom surface 113 can be covered, and the drop height is preferably 0.35 mm or more, more preferably 0.5 mm or more. Therefore, the area of the accommodation space bottom surface 113 is preferably 0.75 mm ² or more, more preferably 3 mm ² or more, further preferably 9 mm ² or more, preferably 80 mm ² or less, more preferably 20 mm ² or less, still more preferably. 16 mm ² or less. By setting the area of the accommodation space bottom surface 113 to 0.75 mm ² or more, a plurality of cells, preferably human fertilized eggs, can be placed and cultured on the accommodation space bottom surface 113 without overlapping each other. By culturing a plurality of cells, particularly fertilized eggs, in the same system, a paracrine effect can be expected in which cell secretions such as proteins, hormones and enzymes accumulated in the culture medium act on each other's cells. Moreover, since it is difficult to evaluate cells and fertilized eggs with a microscope or the like when the cells are overlapped on a plane, the bottom surface preferably has a certain area. Moreover, also when forming the hollow for positioning a cell in the accommodation space bottom face 113 like Embodiment 6 mentioned later, it is preferable that a bottom face has a certain amount of area. Further, by setting the area of the bottom surface 113 of the storage space to 80 mm ² or less, it is possible to cover the entire bottom surface even when a small volume of culture solution is dropped, that is, the drop moves greatly on the bottom surface 113 of the storage space. Can be prevented. The maximum width of the bottom surface 113 of the accommodation space (in the figure in plan view of the outline of the bottom surface, the maximum value of the distance between a pair of points on the periphery of the figure facing the center of gravity of the figure in between, When the bottom surface is a circle, the diameter is preferably 1 mm or more, more preferably 2 mm or more, still more preferably 3.5 mm or more, preferably 10 mm or less, more preferably 5 mm or less, and even more preferably 4.5 mm or less. is there.

  The maximum volume of the liquid that can be stored in the storage space 140 is, for example, 100 μl to 3 ml, particularly 200 μl to 1 ml. By making this volume below a certain level, an autocrine effect due to cell secretions and a paracrine effect due to the interaction of a plurality of cells can be expected. In addition, for example, when fertilized eggs are cultured, a culture technique may be performed in which, for example, the culture medium is replaced with a new culture medium by half after the culture for two days. In these operations, the operation is facilitated by having a culture solution of 200 μl or more.

  The material of the cell culture container 100 of the present invention is not particularly limited. Specifically, inorganic materials such as metal, glass, and silicon, plastics (for example, polystyrene resin, polyethylene resin, polypropylene resin, ABS resin, nylon, acrylic resin, fluororesin, polycarbonate resin, polyurethane resin, methylpentene resin, And organic materials represented by phenol resin, melamine resin, epoxy resin, and vinyl chloride resin). The cell culture container 100 can be manufactured by a method known to those skilled in the art. For example, when a culture container made of a plastic material is manufactured, it can be manufactured by a conventional molding method such as injection molding.

  For observation with an optical microscope, the cell culture vessel 100 is preferably formed of a light transmissive material.

  The cell culture container 100 of the present invention may be surface-treated or surface-coated so as to promote the development of a fertilized egg. In particular, when co-cultured with cells of other organs (for example, endometrial cells or fallopian tube epithelial cells) in order to promote the development of a fertilized egg, these cells are stored in advance in the accommodating space 140 of the cell culture container 100. It is necessary to adhere to the surfaces 113 and 123 facing the surface. In such a case, it is advantageous to coat the surface 113, 123 of the cell culture vessel 100 with a cell adhesive material.

  The cells cultured using the cell culture vessel 100 of the present invention and observed as necessary are not particularly limited. For example, fertilized eggs, egg cells, ES cells (embryonic stem cells) and iPS cells (artificial pluripotency) Stem cells). An egg cell refers to an unfertilized egg cell, and includes an immature oocyte and a mature oocyte. After fertilization, the fertilized egg increases in number of cells at the 2 cell stage, 4 cell stage, and 8 cell stage by cleavage, and develops into a blastocyst through a morula. Fertilized eggs include early embryos such as 2-cell embryos, 4-cell embryos and 8-cell embryos, morulas, blastocysts (including early blastocysts, expanded blastocysts and escaped blastocysts). A blastocyst means an embryo composed of external cells with the potential to form the placenta and internal cell masses with the potential to form embryos. ES cells refer to undifferentiated pluripotent or totipotent cells obtained from the inner cell mass of a blastocyst. An iPS cell refers to a cell having pluripotency similar to that of an ES cell by introducing several types of genes (transcription factors) into somatic cells (mainly fibroblasts). That is, in the present invention, the cells to be cultured include a collection of a plurality of cells such as fertilized eggs and blastocysts. The cell culture vessel 100 of the present invention is suitable for culturing mammalian and avian cells, particularly mammalian cells. Mammals refer to warm-blooded vertebrates such as primates such as humans and monkeys, rodents such as mice, rats and rabbits, pets such as dogs and cats, and domestic animals such as cows, horses and pigs. Can be mentioned. The cell culture vessel 100 of the present invention is particularly suitable for culturing bovine fertilized eggs.

  Hereinafter, other embodiments shown in the drawings will be described, but the scope of the present invention is not limited to these embodiments. In the drawings of the present specification, configurations having the same functions are denoted by the same reference numerals regardless of the dimensions and shapes of the respective configurations, and the description thereof is omitted except for particular differences from the first embodiment.

<Embodiment 2 (FIG. 2)>
FIG. 2 shows Embodiment 2 of the cell culture container 100 of the present invention.
Unlike the first embodiment, the cell culture container 100 according to the second embodiment does not include the outer peripheral wall portion 130 and the outer edge bottom portion 112, and the peripheral wall portion 120 itself standing up from the container bottom portion 110 constitutes the outer peripheral wall portion of the container. . In the second embodiment, the container bottom portion 110 includes an accommodation space bottom portion 111.

  In the second embodiment shown in FIG. 2, one stepped portion 125 that rises in a step shape is formed in a portion other than the vicinity of the opening side end of the outer wall surface 124, but the stepped portion 125 is an essential configuration. Absent.

  In the second embodiment shown in FIG. 2, the outer wall 122 has the same form and dimensions as the outer peripheral wall 130 in the first embodiment, and the inner wall 121 extends to the opening side end of the outer peripheral wall 130 in the first embodiment. It is an extended form. In the second embodiment, the shape of the line 150 in plan view can be an arbitrary shape such as a circular shape (including a circular shape and an elliptical shape), and is preferably a circular shape, and its opening width (in the figure) , The maximum value of the distance between a pair of points on the periphery of the figure facing each other through the center of gravity of the figure) is preferably 30 to 60 mm, and the capacity of the accommodation space 140 is 500 μl to 10 ml is there. Features such as structure, shape, size, angle, and the like of other portions in the second embodiment can be the same as those described above with respect to the first embodiment. Alternatively, as a modification of the second embodiment, a modification in which the size, shape, angle, and capacity of the accommodation space 140 of the peripheral wall 120 (including the inner wall 121 and the outer wall 122) in the second embodiment are the same as those of the first embodiment. That is, a modification in which the outer edge bottom portion 112 and the outer peripheral wall portion 130 do not exist in the first embodiment can be given.

In the second embodiment, the same effect as described above for the first embodiment can be realized.
Features such as structure, shape, size, angle, and the like of other portions in the second embodiment can be the same as those described above with respect to the first embodiment.

<Embodiment 3 (FIG. 3)>
FIG. 3 shows Embodiment 3 of the cell culture container 100 of the present invention.
The third embodiment is an example in which the inclination angle of the inner wall surface 123 is larger than the inclination angle of the outer wall surface 124 at least at the opening side end and in the vicinity thereof, and is an example included in the first embodiment.

  In Embodiment 3, the inclination angle at the opening side end of the inner wall surface 123 and in the vicinity thereof is preferably 30 ° or more, more preferably 50 ° or more, preferably 90 ° or less, more preferably 85 ° or less, and particularly preferably 80 °. The inclination angle at the opening side end of the outer wall surface 124 and in the vicinity thereof is preferably 10 ° or more, more preferably 20 ° or more, particularly preferably 30 ° or more, preferably 70 ° or less, more preferably 60 °. Hereinafter, it is particularly preferably 50 ° or less. In the third embodiment, more preferably, the inclination angles of the inner wall surface 123 and the outer wall surface 124 are at the opening side end of the inner wall surface 123 and the outer wall surface 124 and in the vicinity thereof from the bottom end to the opening end. The inclination angle of the inner wall surface 123 is larger than the inclination angle of the outer wall surface 124.

By forming the inner wall surface 123 as a relatively large inclined surface as in the third embodiment, when bubbles are included in the liquid stored in the storage space 140, the bubbles are likely to gather near the inner wall surface 123. There is. The mechanism is as follows. 11 along the time containing the culture medium D in the housing space 140 of the cell culture vessel 100 of the third embodiment, a portion where the edge is in contact with the inner wall surface 123 of the liquid surface D _S of the culture liquid D in the vertical plane It is the schematic diagram of the end surface cut off. H represents a horizontal surface, E is, indicates the point of contact with the inner wall surface 123 and the liquid surface D _S of the culture medium D, theta ₃ shows the contact angle to the culture solution D of the inner wall surface 123. If the inclination angle theta ₁ of the inner wall surface 123 is greater than the theta _3, the liquid surface D _S of the culture medium D is high at the contact point E, becomes lower as approaching inwardly in the housing space 140 from the contact E. When bubbles are included in the culture medium D stored in the storage space 140, the bubbles are likely to gather near the contact point E, and the central portion of the liquid level of the culture medium D can be easily observed.

<Embodiment 4 (FIG. 4) and Modifications 1 and 2 of Embodiment 4 (FIGS. 12 and 13)>
FIG. 4 shows Embodiment 4 of the cell culture container 100 of the present invention.
In the fourth embodiment, the inner wall surface 123 includes a bottom side region 123 (1) and an opening side region 123 (2). The opening side region 123 (2) of the inner wall surface 123 includes a curved surface formed such that the inclination angle decreases as it approaches the opening side end, and is 0 ° at the opening side end. On the other hand, the outer wall surface 124 is also configured by a bottom side region 124 (1) and an opening side region 124 (2). The opening side region 124 (2) of the outer wall surface 124 includes a curved surface formed such that the inclination angle decreases as it approaches the opening side end and is 0 ° at the opening side end. The inner wall surface 123 and the outer wall surface 124 intersect at the opening side end where both the inclination angles are 0 °, and the surface of the opening side end of the peripheral wall portion 120 is in a state where the cell culture vessel 100 is placed on a horizontal plane. A curved surface having a continuous change in inclination is formed without a bent portion in a cross section of the virtual plane passing through the center of gravity of the accommodation space 140 when viewed in plan. In this case, a portion where the inner wall surface 123 and the outer wall surface 124 intersect does not form a clear line in plan view. In the fourth embodiment, the rounded surface of the intersection of the inner wall surface 123 and the outer wall surface 124 has a radius of curvature of preferably 100 μm or less, more preferably 50 μm or less, and particularly preferably 10 μm or less in the cross section of the virtual plane. The surface can be approximated to a part of the arc. Also in the fourth embodiment, since a flat surface perpendicular to the vertical direction is not formed at the opening side end of the peripheral wall portion 120, it is preferable that cells, foreign substances, and the like are not easily held at the opening side end of the peripheral wall portion 120 during cell culture.

  As a first modification of the fourth embodiment, in the illustrated fourth embodiment, the opening angle 123 (2) of the inner wall surface 123 and the opening side region 124 (2) of the outer wall surface 124 are both closer to the opening side end. However, as shown in FIG. 12, the inclination angle of at least one of the opening-side region 123 (2) of the inner wall surface 123 and the opening-side region 124 (2) of the outer wall surface 124, preferably both opening-side ends, is reduced. This is an example larger than 0 °. In the first modification of the fourth embodiment, the surface at the opening side end of the peripheral wall portion 120 is a curved surface having a bending in which the inclination changes discontinuously at the boundary between the inner wall surface 123 and the outer wall surface 124. Forms a continuous line 150 surrounding the opening of the accommodation space 140 in plan view.

  As a second modification of the fourth embodiment, in the illustrated fourth embodiment, as shown in FIG. 13, the opening side region 123 (2) of the inner wall surface 123 and the opening side region 124 (2) of the outer wall surface 124 are respectively shown in FIG. This is an example extending to the opening side end with a substantially constant inclination. In the second modification of the fourth embodiment, a portion where the inner wall surface 123 and the outer wall surface 124 intersect forms a continuous line 150 that surrounds the opening of the accommodation space 140 in plan view.

  In the modified examples 1 and 2 of the fourth embodiment, the opening side end of the peripheral wall 120 has a sharp shape as in the first embodiment and the like, so that cells, foreign substances, etc. are opened on the opening side of the peripheral wall 120 during cell culture. This is preferable because the possibility of being held on the edge is reduced.

  The inclination angle of the opening side region 123 (2) of the inner wall surface 123 is as described in the range of the inclination angle at the opening side end of the inner wall surface 123 and in the vicinity thereof in <About the inclination angle>, and preferably 10 ° or more. More preferably, it is 20 ° or more, particularly preferably 30 ° or more, preferably 70 ° or less, more preferably 60 ° or less, and particularly preferably 50 ° or less. When the inclination angle of the opening side region 123 (2) of the inner wall surface 123 is equal to or larger than the above lower limit values, it is advantageous because the object is difficult to stay and easily falls on the bottom surface 113 of the accommodation space. This is advantageous because the height of the peripheral wall 120 can be reduced.

  The inclination angle of the bottom side region 123 (1) of the inner wall surface 123 is as described in the range of the inclination angle in the portion closer to the bottom side end than the vicinity of the opening side end of the inner wall surface 123 in <Inclination angle>. Yes, preferably 30 ° or more, more preferably 50 ° or more, particularly preferably 70 ° or more, preferably 90 ° or less, more preferably 85 ° or less, particularly preferably 80 °. It is as follows. More preferably, the inclination angle in the bottom region 123 (1) of the inner wall surface 123 is substantially constant. When the inclination angle of the bottom region 123 (1) of the inner wall surface 123 is equal to or larger than the above lower limit values, it is advantageous because the object is difficult to stay and easily falls on the bottom surface 113 of the accommodation space. Since the storage space 140 becomes wider as it approaches the upper side, the capacity of the storage space 140 can be increased, which is advantageous.

  In a more preferable embodiment of Embodiment 4 and Modifications 1 and 2, when the width along the vertical direction from the bottom side end to the opening side end of the inner wall surface 123 is 100, from the bottom side end of the inner wall surface 123. The portion of the bottom region 123 (1) is preferably 50 or more, more preferably 60 or more, particularly preferably 70 or more, preferably 90 or less, more preferably 80 or less along the vertical direction toward the opening side end. . Further, when the width along the vertical direction from the bottom side end to the opening side end of the inner wall surface 123 is 100, it is preferably 50 along the vertical direction from the opening side end of the inner wall surface 123 toward the bottom side end. Less than, more preferably less than 40, particularly preferably less than 40, preferably more than 10, more preferably more than 20, is the opening-side region 123 (2). When the proportion of the width of the bottom side region 123 (1) in the width of the inner wall surface 123 is larger than the proportion of the width of the opening side region 123 (2), the height of the bottom side region 123 (1) ( By adjusting the (vertical width), it becomes easy to design the volume of the accommodation space 140 to a desired volume.

  The inclination angle of the opening-side region 124 (2) of the outer wall surface 124 is as described as the range of the inclination angle at the opening-side end of the outer wall surface 124 and the vicinity thereof in <Regarding the inclination angle>, preferably 10 ° or more. More preferably, it is 20 ° or more, particularly preferably 30 ° or more, preferably 70 ° or less, more preferably 60 ° or less, and particularly preferably 50 ° or less. When the inclination angle of the opening side region 124 (2) of the outer wall surface 124 is equal to or larger than the above lower limit values, it is advantageous because the object is difficult to stay and easily falls to the bottom of the outer peripheral space 160, and is less than the upper limit values described above. This is advantageous because the height of the peripheral wall 120 can be reduced.

  The inclination angle of the bottom side region 124 (1) of the outer wall surface 124 is as described in the range of the inclination angle in the portion closer to the bottom end than in the vicinity of the opening end of the outer wall 124 in <Inclination angle>. Yes, preferably 30 ° or more, more preferably 50 ° or more, particularly preferably 70 ° or more, preferably 90 ° or less, more preferably 85 ° or less, particularly preferably 80 °. It is as follows. More preferably, the inclination angle in the bottom region 124 (1) of the outer wall surface 124 is substantially constant. When the inclination angle of the bottom region 124 (1) of the outer wall surface 124 is equal to or greater than the above lower limit values, it is advantageous because the object is difficult to stay and easily falls to the bottom of the outer peripheral space 160. In this case, when the cell culture container 100 of the present invention is manufactured by injection molding, it is advantageous because it is easy to manufacture because there is a taper, and since the outer peripheral space 160 becomes wider as it approaches the upper side, Advantageously, the volume can be increased.

  In a more preferred embodiment of Embodiment 4 and Modifications 1 and 2, when the width along the vertical direction from the bottom side end to the opening side end of the outer wall surface 124 is 100, from the bottom side end of the outer wall surface 124. The bottom region 124 (1) is preferably 50 or more, more preferably 60 or more, particularly preferably 70 or more, preferably 90 or less, more preferably 80 or less along the vertical direction toward the opening side end. . Further, when the width along the vertical direction from the bottom side end to the opening side end of the outer wall surface 124 is 100, it is preferably 50 along the vertical direction from the opening side end to the bottom side end of the outer wall surface 124. A portion of less than, more preferably less than 40, particularly preferably less than 30, preferably more than 10, more preferably more than 20, is the open side region 124 (2). Thus, when the ratio of the bottom side region 124 (1) of the outer wall surface 124 is larger than the opening side region 124 (2), by adjusting the height (vertical width) of the bottom side region 124 (1), It becomes easy to design the volume of the outer peripheral space 160 to a desired volume.

  Features such as structures, shapes, dimensions, and angles of other portions in the fourth embodiment and the first and second modifications can be the same as those described above with respect to the first embodiment.

<Embodiment 5 (FIG. 5)>
FIG. 5 shows a fifth embodiment of the cell culture container 100 of the present invention.
In the cell culture container 100 of the fifth embodiment, the outer wall surface 124 of the peripheral wall 120 includes at least a bottom region 124 (3) having an inclination angle larger than 90 °, and the inner wall surface 123 and the outer wall surface 124 are open. It intersects at the side end. The cell culture container 100 of Embodiment 5 also includes an outer edge bottom portion 112 projecting outward from the bottom side end of the peripheral wall portion 120, and an outer peripheral wall portion 130 erected on the periphery of the outer edge bottom portion 112. An outer peripheral space 160 surrounded by the outer edge bottom portion 112 and the peripheral wall portion 120 and opened upward is formed.

  In the cell culture using the cell culture vessel 100 including the outer peripheral wall portion 130 and the outer edge bottom portion 112 as in the first, third, and fourth embodiments, the outer peripheral wall portion 130, the outer edge bottom portion 112, and the peripheral wall portion 120 are surrounded. There is a case where a liquid different from the accommodation space 140 is accommodated in the outer peripheral space 160 and cultured. In this usage, as shown in FIGS. 1A and 1B, the inclination angle of the outer wall surface 124 of the peripheral wall 120 is substantially constant at an angle close to 90 ° from the bottom side end to the opening side end. As shown in the drawing, when the inclination angle of the outer wall surface 124 of the peripheral wall portion 120 is substantially constant at an angle of less than 90 ° from the bottom side end to the opening side end, or as in the fourth embodiment illustrated in FIG. When the inclination angle of the outer wall surface 124 is a combination of a portion having an angle close to 90 ° and a portion having an angle of less than 90 °, the liquid stored in the outer circumferential space 160 due to vibration during use is stored in the storage space 140. Easy to flow in.

  As a means for solving the above problems, a cell culture container 600 of a comparative example shown in FIG. 6 can be considered. In the cell culture container 600, the peripheral wall 120 in the first embodiment or the like is replaced with the peripheral wall 610. The peripheral wall portion 610 includes an inner wall portion 121, an outer wall portion 122, and an upper wall portion 602 that connects the inner wall portion 121 and the opening side end of the outer wall portion 122, and the outer wall surface 124 that faces the outer peripheral space 160. The tilt angle is greater than 90 °. According to this configuration, when the liquid is accommodated in the outer circumferential space 160 so that the liquid level is positioned below the opening side end of the outer wall surface 124, the liquid in the outer circumferential space 160 may be subjected to vibration or the like during culture. There is an advantage that it is difficult for the liquid to flow into the storage space 140. However, in the cell culture container 600 of the comparative example shown in FIG. 6, the opening side ends of the inner wall surface 123 and the outer wall surface 124 do not intersect with each other and are connected by the upper wall portion 602 having an inclination angle of approximately 0 °. Therefore, there is a problem that cells, foreign substances, and the like are easily held on the upper wall portion 602.

  The fifth embodiment shown in FIG. 5 is intended to solve the above problem. That is, the inclination angle of the bottom region 124 (3) of the wall surface 124 is greater than 90 °, preferably 95 ° or more, more preferably 100 ° or more, preferably 140 ° or less, more preferably When the liquid is accommodated in the outer peripheral space 160 so that the liquid level is positioned below the opening side end of the bottom side region 124 (3) of the outer wall surface 124 by being 120 ° or less, Even in the case of vibration or the like, the liquid in the outer peripheral space 160 hardly flows into the storage space 140. In addition, due to the configuration in which the inner wall surface 123 and the outer wall surface 124 intersect at the opening side end, cells, foreign substances, and the like are hardly held at the opening side end of the peripheral wall portion 120 as in the first embodiment.

  When the width along the vertical direction from the bottom side end to the opening side end of the outer wall surface 124 is 100, preferably 50 or more along the vertical direction from the bottom side end of the outer wall surface 124 toward the opening side end. More preferably 60 or more, particularly preferably 70 or more, preferably 90 or less, more preferably 80 or less is the bottom region 124 (3) having the above-mentioned inclination angle. More preferably, the bottom region 124 (3) is constituted by a surface having a substantially constant inclination angle within the above range.

  The inclination angle in the vicinity of the opening side end of the outer wall surface 124 is as described in the first embodiment. More preferably, the opening side region 124 (4) of the outer wall surface 124 is a region having a substantially constant inclination angle of 10 ° or more and less than 90 °, preferably 30 or more and 70 ° or less. When the width along the vertical direction from the bottom side end to the opening side end of the outer wall surface 124 is 100, preferably less than 50 along the vertical direction from the opening side end to the bottom side end of the outer wall surface 124, More preferably, the portion of less than 40, particularly preferably less than 30, preferably more than 10, more preferably more than 20, is the opening side region 124 (4) having the above inclination angle.

  On the other hand, the inclination angle of the inner wall surface 123 can be set as appropriate so that it can intersect the outer wall surface 124 at the opening side end, and the preferable range of the inclination angle is as described in <Inclination angle>. More preferably, as shown in FIG. 5, the inner wall surface 123 includes a bottom region 123 (3) and an opening region 123 (4).

  The inclination angle of the bottom side region 123 (3) of the inner wall surface 123 is preferably larger than 0, more preferably 15 ° or more, preferably 90 ° or less, more preferably 80 ° or less. More preferably, the bottom side region 123 (3) of the inner wall surface 123 is configured by a surface having a substantially constant inclination angle within the above range. When the width along the vertical direction from the bottom side end to the opening side end of the inner wall surface 123 is 100, preferably 50 or more along the vertical direction from the bottom side end to the opening side end of the inner wall surface 123, More preferably 60 or more, particularly preferably 70 or more, preferably 90 or less, more preferably 80 or less is the bottom region 123 (3) having the above-mentioned inclination angle.

  The inclination angle of the opening-side region 123 (4) of the inner wall surface 123 is preferably 10 ° or more, more preferably 30 ° or more, preferably 90 ° or less, more preferably 70 ° or less. The opening side region 123 (4) of the inner wall surface 123 is more preferably configured by a surface having a substantially constant inclination angle within the above range. When the width along the vertical direction from the bottom side end to the opening side end of the inner wall surface 123 is 100, preferably less than 50 along the vertical direction from the opening side end to the bottom side end of the inner wall surface 123, More preferably, the portion of less than 40, particularly preferably less than 30, preferably more than 10, more preferably more than 20, is the opening side region 123 (4) having the above inclination angle.

  Features such as structure, shape, size, angle, and the like of other parts in the fifth embodiment can be the same as those described above with respect to the first embodiment.

<Embodiment 6 (FIGS. 7A and 7B)>
In the cell culture container 100 of the present invention, it is further preferable that a recess 114 for positioning cells is formed on the bottom surface 113 of the storage space, which is a surface forming the bottom of the storage space 140. This embodiment of the present invention is shown as Embodiment 6 in FIGS. 7A and 7B.

  The cell culture container 100 according to Embodiment 6 shown in FIGS. 7A and 7B has the same configuration as that of the cell culture container 100 according to Embodiment 1 except that a plurality of depressions 114 are formed on the bottom surface 113 of the accommodation space. Prepare. By disposing the cells in the recess 114, the movement of the cells can be suppressed, and the evaluation and determination can be performed after each cell is specified. The shape of the recess is not particularly limited, but it is preferable that the outer edge of the opening is circular and the wall surface of the recess has a curved surface that is inclined so as to increase from the lowest position toward the outer edge. The wall surface of the depression preferably includes a conical or frustoconical portion. The conical or frustoconical portion is formed on the bottom side of the culture vessel so that the apex of the cone or the upper surface and the lower surface of the truncated cone has the smaller area. Conical shapes include cones and elliptical cones, and similar shapes, such as cones or elliptical cones with rounded vertices, conical surfaces bulging outwards, conical surfaces recessed inwards Shape etc. are included. The truncated cone has a truncated cone and an elliptical frustum, and similar shapes, for example, a shape in which the junction between the upper or lower surface of the truncated cone or the truncated truncated cone and the truncated cone is rounded, and the truncated cone is on the outer side. And a shape having a conical surface recessed inward. The depression 114 is not limited to the above, but may have a polygonal pyramid shape, a polygonal frustum shape, or the like.

  The size of the recess 114 is not particularly limited as long as it is a size that can accommodate at least one cell. Here, the dimension of the depression refers to the length of the longest diameter of the figure formed by the outer edge of the opening of the depression 114. Therefore, when the outer edge of the opening of the depression 114 is circular, the diameter is the same as or larger than the maximum dimension of the cells to be cultured. When fertilized eggs are cultured in the cell culture vessel of the present invention, it is desirable to culture to the blastocyst stage, so the diameter of the circular opening is larger than the maximum cell size of the blastocyst stage It is desirable. Since the maximum size of cells in the blastocyst stage is usually 100 μm to 280 μm, the diameter of the circular opening is usually 100 μm or more.

  For example, in the case of a human fertilized egg, the diameter of the opening of the depression 114 is usually 100 μm or more, preferably 200 μm or more, more preferably 250 μm or more, and usually 1000 μm or less, preferably 900 μm or less, more preferably 800 μm or less. . Moreover, the diameter of the opening part of the hollow 114 can also be prescribed | regulated as X + (alpha) (here X represents the maximum dimension of a cell). Here, α is preferably 0.01 mm or more, and more preferably 0.02 mm or more.

  Features such as structure, shape, size, angle, and the like of other portions in the sixth embodiment can be the same as those described above with respect to the first embodiment.

100: cell culture container 140: accommodation space 110: container bottom 120: peripheral wall portion 123: inner wall surface 124: outer wall surface 150: a continuous line surrounding the opening of the accommodation space 140 in plan view

Claims (2)

A cell culture container in which an accommodation space for accommodating cells and a culture solution is formed,
The housing space is a space that is surrounded by a peripheral wall portion that stands up from the bottom of the container and opens upward.
The peripheral wall portion has an inner wall surface facing the housing space and an outer wall surface facing the outer side of the housing space,
The cell culture container, wherein the inner wall surface and the outer wall surface intersect at an opening side end.   The cell culture container according to claim 1, wherein a portion where the inner wall surface intersects with the outer wall surface is a continuous line surrounding the opening of the accommodation space in plan view.

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