TW201623604A - Culture container - Google Patents

Abstract

A culture container includes a bottom surface and a culture chamber including a peripheral sidewall surface which is provided around a periphery of the bottom surface, wherein the culture chamber is provided with a barrier surface having a predetermined height around the periphery and the barrier surface extends vertically and toward the peripheral sidewall surface from an inner periphery on the bottom surface, which is inside the periphery, as a base end and is formed so as to make a border line between the peripheral sidewall surface and the barrier surface.

Description

Culture container

The present invention relates to a culture vessel. More specifically, the present invention relates to a culture vessel having a meniscus resistant effect. The present application is based on Japanese Patent Application No. 2014-195164, filed on Jan.

When the culture solution is added to the culture vessel, a concave meniscus is generated at the interface due to the interfacial tension of the solution. In particular, in the case where a gel is used for the culture, the tendency becomes more conspicuous because the viscosity of the solution before gelation is high and the solution tends to adhere to the inner wall surface of the culture vessel. When the culture surface is observed, a shadow is formed in the portion where the meniscus is formed, thereby obstructing the observation of the cultured cells existing in the portion.

As a technique for solving the problem of the meniscus, a cell culture device is disclosed in Japanese Laid-Open Patent Publication No. 2001-340070 (Patent Document 1), which has a layer A on the bottom surface of the culture container and on the layer A. Superposedly having a layer B containing a substance derived from a living body, and a solution which becomes a gel state by heating is frozen in an ungelled state, and the layer B is formed into a gel by heating. The aqueous solution of the state is frozen.

[Patent Document 1] Japanese Patent Laid-Open Publication No. 2001-340070

The conventional technique for solving the problem of the meniscus is accomplished by "acting to reduce or prevent the occurrence of the meniscus itself".

On the other hand, an object of the present invention is to provide a culture container which allows the generation of a meniscus and which solves the problem of the meniscus.

As a result of intensive studies, the inventors of the present invention have achieved the object of the present invention by providing a structure in which the culture vessel has "the meniscus is disposed outside the range to be cultured and observed", and completed the present invention.

(1)

The culture container of the present invention comprises a culture chamber having a bottom surface and a circumferential side wall surface provided on a periphery of the bottom surface. In the culture chamber, a barrier surface is provided around the circumference of the bottom surface. The barrier surface is formed such that the inner peripheral edge of the bottom surface on the inner side of the outer periphery rises as a base end, faces the peripheral side wall surface, and forms a boundary with the peripheral side wall surface.

The inner peripheral edge is formed at a position spaced apart from the inner side of the circumference to at least reduce the influence of the meniscus generated when the culture liquid is accommodated in the culture chamber. The barrier surface is set at a height that can form a degree of difference: the degree of cell residue on the barrier surface is suppressed.

According to the above configuration, it is possible to reduce the cultivation which is difficult to observe due to the occurrence of the meniscus on the culture liquid surface. The cultured cells are present in the peripheral part of the nursery. Therefore, although the meniscus is allowed to be produced, the observation range of the cultured cells can be expanded.

(2)

Alternatively, the shape of the barrier surface may be a straight line, a curved line, or a combination of a straight line and a curved line from a transverse cross-sectional view.

In this way, the barrier faces of various shapes are allowed to widen.

(3)

Preferably, since the transverse cross-sectional view, the barrier surface is formed by a lower barrier surface portion that rises from the inner peripheral edge and an upper barrier surface portion that is connected to the lower barrier surface portion and forms a boundary with the peripheral sidewall surface, and rises from the inner peripheral edge. The lower side barrier surface and the bottom surface are at an angle of 90 degrees or more and 150 degrees or less.

When the angle of the lower barrier surface portion is 90 degrees or more, the observation range of the concentrated cultured cells can be expanded without obstructing the field of view of the portion other than the peripheral portion in the culture chamber, and the culture can be further reduced by 150 degrees or less. The cells enter the peripheral portion of the culture chamber that is difficult to observe.

(4)

In the culture container according to the above (3), the side barrier portion and the upper barrier portion may be connected to each other without a boundary from the same slope in the transverse cross-sectional view.

In this case, the structure of the barrier surface is simple, and it is also preferable from the viewpoint of manufacturing.

(5)

When the shape of the barrier surface is a curve from the transverse cross section, the curve preferably has the following shape: The angle formed by the line from the bottom surface is increased from the bottom surface side toward the peripheral side wall surface side. That is, it may be a shape in which the shape of the barrier surface is a curve when viewed from the lateral direction (the direction parallel to the bottom surface), and the curve is such that the angle formed by the tangent to the bottom surface gradually changes from the bottom surface side toward the peripheral side wall surface side. Big shape.

In this case, it is preferable to ensure that the cultured cells are not present in the peripheral portion of the culture chamber which is difficult to observe, and it is also preferable from the viewpoint of production.

(6)

The shape of the circumferential side wall surface and the inner circumference of the circumference may be a circle. In this case, the diameter of the inner circumference may be 30 mm or more and 170 mm or less.

By having a diameter of 30 mm or more in the inner circumference, it is preferable to obtain a space on the bottom surface for cultivation and observation, and by being 170 mm or less, it is preferable to concentrate most of the cultured cells. The effect of the observed part.

According to the present invention, it is possible to provide a culture container in which the shadow formed by the meniscus does not hinder the observation of the cultured cells even if a meniscus is generated.

100, 1001‧‧‧ culture container

200‧‧‧ bottom

300, 300b, 300d‧‧‧ weeks side wall

400, 400a, 400b, 400c, 400d, 400e, 400f, 400g, 400h, 400i, 400j, 400k‧ ‧ barrier

410, 410f, 410g, 410h, 410j, 410k‧‧‧ lower barrier face

420, 420d, 420e, 420f, 420g, 420h, 420j, 420k‧‧‧ upper barrier face

O, B‧‧‧ (bottom) circumference

B2, B2a‧‧‧ (bottom) internal circumference

B3, B3b, B3d‧‧‧ (Bound wall and circumferential side wall) boundary

C‧‧‧Cultivation room

θ 1‧‧‧ (the lower barrier face and the bottom surface)

θ, θ', θ"‧‧‧ (the tangent to the bottom of the cross-sectional shape of the barrier surface)

Fig. 1 is a schematic external perspective view of a culture container according to a first embodiment.

Fig. 2 is a schematic cross-sectional view showing the use of the culture container of the first embodiment.

Fig. 3 is a partial enlarged view of the schematic sectional view of Fig. 2;

Fig. 4 is an enlarged view of an essential part of the schematic sectional view of Fig. 2;

Fig. 5 is a schematic cross-sectional view showing the use of the culture container according to the first modification of the first embodiment.

Fig. 6 is a schematic cross-sectional view showing the use of the culture container according to the second modification of the first embodiment.

Fig. 7 is a schematic cross-sectional view showing the use of the culture container according to the third modification of the first embodiment.

Fig. 8 is a schematic cross-sectional view showing the use of the culture container according to the fourth modification of the first embodiment.

Fig. 9 is a schematic cross-sectional view showing the use of the culture container according to the fifth modification of the first embodiment.

Fig. 10 is an enlarged view of a main part of a culture container according to a second embodiment.

Fig. 11 is an enlarged view of a main part of a culture container according to a third embodiment.

Fig. 12 is an enlarged view of a main part of a culture container according to a fourth embodiment.

Fig. 13 is an enlarged view of a main part of a culture container according to a fifth embodiment.

Fig. 14 is an enlarged view of a main part of the culture container according to the sixth embodiment.

Fig. 15 is an enlarged view of a main part of a culture container according to a seventh embodiment.

Fig. 16 is a schematic perspective view showing the culture container of the eighth embodiment.

Fig. 17 is a photograph of the culture container prepared in Example 1 after being added to the culture medium, and then viewed from above and below.

Fig. 18 is a photograph (partial enlargement) of the cell culture after the cell culture was carried out in the culture vessel prepared in Example 1.

Figure 19 is a view of the up-and-down direction after cell culture in the culture vessel prepared in Example 1. Photo (partial enlargement).

Fig. 20 is a photograph of the culture container prepared in Example 2, which was observed in the vertical direction after being added to the culture medium.

Fig. 21 is a photograph (partial enlargement) of the observation in the lower direction after cell culture in the culture vessel prepared in Example 2.

Fig. 22 is a photograph (partial enlargement) of the cell culture in the culture vessel prepared in Example 2, which was observed in the vertical direction.

Fig. 23 is a schematic cross-sectional view showing a culture vessel prepared in Comparative Example 1.

Fig. 24 is a photograph of the culture container prepared in Comparative Example 1 which was observed in the vertical direction after the medium was added.

Fig. 25 is a photograph (partial enlargement) of the cell culture after the cell culture was carried out in the culture vessel prepared in Comparative Example 1.

Fig. 26 is a photograph (partial enlargement) of the cells cultured in the culture vessel prepared in Comparative Example 1 and observed in the vertical direction.

Hereinafter, embodiments of the present invention will be described with reference to the drawings. In the following description, the same elements are denoted by the same reference numerals, and the names and functions thereof are the same unless otherwise specified. Therefore, the detailed description thereof will not be repeated.

[First Embodiment]

Fig. 1 is a schematic external perspective view of a culture container according to a first embodiment. Fig. 2 is a schematic cross-sectional view showing the use of the culture container of the first embodiment. Figure 2 is a schematic cross-sectional view of the horizontal The view of the surface cut by the vertical plane including the line A-A of Fig. 1 was observed. Fig. 3 is a partial enlarged view of the schematic sectional view of Fig. 2. Fig. 4 is an enlarged view showing an essential part (circular portion) of the schematic sectional view of Fig. 2. In addition, for convenience of description, the upper side of each drawing is called upper, and the lower side is called lower.

As shown in FIGS. 1 and 2, the culture container 100 includes a bottom surface 200, a peripheral side wall surface 300, and a barrier wall surface 400 as surfaces constituting the inner side thereof. The culture chamber C is defined by the bottom surface 200, the peripheral side wall surface 300, and the barrier surface 400. In other words, the culture container according to the present embodiment includes the culture chamber C including the bottom surface 200, the barrier wall surface 400 provided outside the peripheral edge B2 of the bottom surface 200, and the circumference provided on the outer side along the periphery of the barrier surface 400. Side wall surface 300.

The bottom surface 200 is circular in the vertical direction, and as shown in FIG. 3, a peripheral side wall surface 300 which is raised from the entire circumference O of the bottom surface 200 and which is opened upward is provided. The barrier rib 400 disposed around the entire circumference of the peripheral edge O of the bottom surface 200 at a specific height h is formed by raising the inner periphery B2 of the specific distance w from the periphery O of the bottom surface 200 as a base end. The peripheral side wall surface 300 is formed, and a boundary B3 with the peripheral side wall surface 300 is formed at a position of a specific height h.

As shown in FIG. 3, the specific distance w corresponds to the distance between the circumference O and the inner circumference B2. The specific height h corresponds to the distance between the peripheral edge O and the boundary B3, and is set so as to prevent the cell from remaining in the shape of the barrier rib 400 by the relationship with the specific distance w. In the present embodiment, it is set to be equal to or lower than the lowest water level H of the culture solution to be contained in the culture chamber C.

When the meniscus surface M is generated in the culture liquid surface S, the culture liquid surface S of the peripheral portion of the culture chamber C is curved due to the meniscus M phenomenon, so that the field of view viewed from the up and down direction becomes difficult to observe in this portion. . In the culture vessel 100, the cultured cells are prevented from moving from the inner circumference B2 to the barrier surface 400 by providing the barrier surface 400 at a specific height h rising from the inner periphery B2. The side movement is such that at least a part of the portion where the meniscus M is generated is disposed outside the inner circumference B2 by causing the barrier surface 400 to face outward from the inner circumference B2 and the inner circumference B2 and the peripheral side wall surface 300 by a specific distance w. . Therefore, although the meniscus M is allowed to be generated, most of the cultured cells can be present inside the inner periphery B2 which can be observed.

The barrier surface 400 has a width corresponding to the specific distance w, and at least reduces the influence of the meniscus M generated on the culture liquid surface S when the culture liquid is contained in the culture chamber C. Therefore, the specific distance w is not particularly limited. For example, the larger the specific distance w is, the more the tendency of the meniscus M is further reduced, and the smaller the specific distance w, the more the space for the culture and observation of the bottom surface 200 is sufficiently obtained. It is appropriately determined based on these two tendencies. As a specific example, the specific distance w may be 2 mm or more and 3 mm or less. In the present embodiment, the specific distance w is set to be equal to or larger than the width W of the meniscus M from the viewpoint of avoiding the influence of the meniscus M.

The specific height h is not particularly limited as long as it can be set as follows, and the difference is such that the difference in cell retention between the barrier faces 400 is suppressed by the relationship with the specific distance w. For example, the specific height h is 1 mm or more and 21 mm or less, preferably 1 mm or more and 15 mm or less. By setting it to the above lower limit value, it is possible to further reduce the outer portion of the inner peripheral edge B2 which is difficult to be observed when the cultured cells are infiltrated in the vertical direction, and it is possible to sufficiently obtain the culture liquid to be accommodated by being equal to or less than the above upper limit value. Volume. In the present embodiment, the specific height h is set to be equal to or lower than the water level H of the culture solution to be accommodated in the culture chamber C from the viewpoint of more sufficiently acquiring the volume of the culture solution to be stored. In this case, the boundary E between the culture liquid surface S and the inner wall surface of the culture vessel 100 is on the peripheral side wall surface 300.

As shown in FIG. 4, the barrier surface 400 is composed of a lower barrier surface portion 410 and an upper barrier surface. The unit 420 is configured. The lower barrier surface portion 410 is raised from the base end B2 so as to be at an angle θ 1 to the bottom surface 200, and is connected to the upper barrier faucet portion 420 at a position of the height h1. At the position of the height h1, a ridge line is formed at the boundary between the lower barrier portion 410 and the upper barrier portion 420. The upper barrier façade 420 is inclined toward the peripheral side wall surface 300 at an angle θ 2 with respect to the bottom surface 200, and forms a boundary B3 with the circumferential side wall surface 300.

The angle θ 1 formed by the lower barrier surface portion 410 and the bottom surface 200 is 90 degrees. Thereby, it is possible to effectively prevent the cultured cells in the bottom surface 200 from moving outward beyond the inner periphery B2. The angle θ 2 of the inclination angle of the upper barrier surface portion 420 is not particularly limited. For example, there is a tendency that the larger the angle θ 2 is, the more the height h1 of the lower barrier surface portion 410 can be sufficiently ensured, and the tendency of the cultured cells to infiltrate into the outer portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up-and-down direction is further reduced; The smaller the angle θ 2 is, the more the volume of the culture solution to be accommodated can be sufficiently obtained; the operator can appropriately determine the two tendencies based on the two tendencies.

The height h1 of the lower barrier surface portion 410 is not particularly limited. For example, there is a tendency that the larger the height h1 is, the more the "the outer portion of the inner peripheral edge B2 which is difficult to be observed when the cultured cells are infiltrated in the vertical direction" is further reduced; the smaller the height h1, the more the volume of the culture medium to be accommodated can be sufficiently obtained. The tendency of the industry can be appropriately determined based on these two tendencies. Specifically, the height h1 can be set, for example, to 0.5 mm, or can be set to 1 mm.

Diameter of the inner circumference B2 It can be the same diameter as the inner bottom surface of the conventional culture container. For example, it is 30 mm or more and 170 mm or less, preferably 32 mm or more and 90 mm or less. By being at least the above lower limit value, it is possible to preferably obtain a space on the bottom surface 200 for cultivation and observation, and by being equal to or less than the above upper limit value, it is preferable to enjoy the large size of the cultured cells. Partly focused on the effects of the observable parts.

The culture container 100 can be composed of a raw material having transparency. Examples of the transparent material include inorganic materials represented by glass and quartz, and organic materials represented by synthetic resins. Examples of the synthetic resin include polystyrene (PS), polypropylene (PP), polymethylpentene (PMP), polycarbonate (PC), polymethyl methacrylate (PMMA), and polymethyl methacrylate. Polymers such as quinone imine (PMMI) and cycloolefin copolymer (COC). Further, a copolymer synthesized from two or more monomer units of the polymers may also be mentioned. In the case of a synthetic resin, it is preferable in terms of being easily formed and not easily broken.

The culture container 100 may be an integrally formed structure, or may be a structure formed by, for example, a combination of a member constituting the bottom surface 200 and members constituting the peripheral side wall surface 300 and the barrier wall surface 400. In the case of a structure formed by a combination of members, each member may be composed of different raw materials.

The bottom surface 200, the peripheral side wall surface 300, and the barrier wall surface 400 of the culture vessel 100 may also be subjected to a surface treatment in terms of physical, chemical, and/or biochemical viewpoints. As long as it is a manufacturer, such a surface treatment can be appropriately selected. For example, the barrier surface 400, or the barrier surface 400 and the peripheral sidewall surface 300 may be subjected to a surface treatment for hindering cell culture.

Further, a further additional structure (not shown) may be provided depending on the purpose of use of the culture container 100 or the like. As long as it is a manufacturer, such an additional structure can be appropriately selected. Specifically, a lid body for making the culture chamber C into a capped state, a spacer for spacing the contents, and a grid pattern for specifying the amount and position of the contents (particularly, a cell population) may be mentioned. A guiding structure provided on the bottom surface 200.

<Modification>

The culture container 100 is allowed to have various modifications in addition to those already illustrated. In the variant, As long as at least a part of the meniscus M is disposed outside the inner peripheral edge B2, any configuration may be allowed as long as the boundary E is not formed on the lower barrier surface 410 in the barrier face 400, and the lower barrier face 410 may be allowed. The inner circumference B2 is used as a base end and is raised at 90 degrees with respect to the bottom surface 200. In other words, in the modified example, when the culture liquid has been stored in the culture chamber C, the boundary E between the liquid surface S of the culture liquid and the inner wall of the culture container 100 may be configured to be outside the inner circumference B2.

Hereinafter, a modification of the first embodiment in which the combination of the specific distance w and the specific height h is different will be described with reference to FIGS. 5 to 9. 5 to 9 correspond to Fig. 3 of the first embodiment. In the following modifications, the differences from the first embodiment will be mainly described, and the description of the same aspects will be omitted.

[First Modification]

Fig. 5 shows a first modification of the first embodiment. The barrier surface 400a of the culture container 100a shown in Fig. 5 is set to be shorter than the specific distance w of the first embodiment by setting the specific distance wa to be shorter than the inner circumference B2a of the first embodiment. B2 is more lateral. Thereby, the meniscus M provided outside the inner peripheral edge B2a becomes a part of the barrier face 400a, and the influence by the meniscus M can be reduced. That is, in the present modification, since the meniscus M is disposed outside the inner peripheral edge B2a, the influence due to the meniscus M can be reduced.

[Second Modification]

Fig. 6 shows a second modification of the first embodiment. The barrier surface 400b of the culture container 100b shown in Fig. 6 is set to be larger than the specific height h of the first embodiment by the specific height hb, so that the boundary B3b with the peripheral side wall surface 300b is at the boundary B3 of the first embodiment. More on the side. Thereby, the culture cells can be further infiltrated into the inner periphery B2 which is difficult to observe when viewed from the top and bottom direction. In the present modification, the specific height hb is equal to or higher than the water level H of the culture solution to be accommodated in the culture chamber C, but does not reach the height E of the boundary E between the culture liquid surface S and the circumferential side wall surface 300b.

[Third Modification]

Fig. 7 shows a third modification of the first embodiment. In the same manner as in the first modification, the barrier wall surface 400c of the culture container 100c shown in Fig. 7 is such that the base end, that is, the inner circumference B2a, is set by setting the specific distance wa to be shorter than the specific distance w of the first embodiment. The inner periphery B2 of the first embodiment is further outside. Thereby, the meniscus M provided outside the inner peripheral edge B2a becomes a part of the barrier face 400c, and the influence by the meniscus M can be reduced. That is, in the present modification, since the meniscus M is disposed outside the inner peripheral edge B2a, the influence due to the meniscus M can be reduced.

Further, similarly to the second modification, the boundary B3b between the barrier rib surface 400c and the peripheral side wall surface 300b is higher than the boundary B3 of the first embodiment. Thereby, it is possible to further reduce the infiltration of the cultured cells into the "outer side portion of the inner peripheral edge B2a which is difficult to observe when viewed in the up-and-down direction". In the present modification, the specific height hb is equal to or higher than the water level H of the culture solution to be accommodated in the culture chamber C, but does not reach the height E of the boundary E between the culture liquid surface S and the circumferential side wall surface 300b.

[Fourth Modification]

Fig. 8 shows a fourth modification of the first embodiment. The barrier surface 400d of the culture container 100d shown in Fig. 8 is set to be larger than the specific height hb of the second modification by the specific height hd, and the boundary B3d with the peripheral side wall surface 300d is in the second modification. The boundary B3b is further on the upper side. Thereby, it is possible to further reduce the infiltration of the cultured cells into the "outer side portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up-and-down direction".

In the present modification, the culture liquid surface S and the inner wall surface of the culture container 100d are Since the boundary Ed is on the upper peripheral side wall surface portion 420d, the specific height hd of the barrier surface 400d exceeds the water level H of the culture liquid to be accommodated in the culture chamber C and is equal to or higher than the height of the boundary Ed. However, since a sufficient amount of the specific distance w is provided, the meniscus M can be disposed on the outer side of the inner circumference B2, and the influence of the meniscus M can be avoided.

[Fifth Modification]

Fig. 9 shows a fifth modification of the first embodiment. Similarly to the fourth modification, the barrier surface 400e of the culture container 100e shown in FIG. 9 is formed so as to have a specific height hd larger than the specific height hb of the second and third modifications. The boundary B3d of 300d is at a higher boundary than the boundary B3b of the second and third modifications. Thereby, it is possible to further reduce the infiltration of the cultured cells into the "outer side portion of the inner peripheral edge B2a which is difficult to observe when viewed in the up-and-down direction".

In the present modification, the boundary Ee between the culture liquid surface S and the inner wall surface of the culture container 100e is on the upper circumferential side wall surface portion 420e. Therefore, the specific height hd of the barrier surface 400e exceeds the water level H of the culture liquid to be accommodated in the culture chamber C. And it is above the height of the boundary Ee. Further, similarly to the first modification, by setting the specific distance wa to be shorter than the specific distance w of the first embodiment, the inner end B2a which is the base end is located outside the inner periphery B2 of the first embodiment. Thereby, the meniscus M provided on the outer side of the inner peripheral edge B2a becomes a part of the barrier face 400e, and the influence by the meniscus M can be reduced. That is, in the present modification, since the meniscus M is provided outside the inner peripheral edge B2a, the influence by the meniscus M can be reduced.

<Other Embodiments>

In the present invention, the barrier surface 400 is not limited to the first embodiment as long as it has a shape passing between the inner peripheral edge B2 and the boundary B3 (a shape connecting the inner peripheral edge B2 and the boundary B3) from the transverse cross-sectional view. The shape. For example, the lower barrier portion 410 and the upper barrier portion 420 The boundary may be formed as shown in FIG. 4, or C-chamfering or R-chamfering may be performed on the boundary.

Further, the cross-sectional shape of the barrier surface 400 from the lateral direction may be a straight line, a curved line, or a combination of a straight line and a curved line, and various shapes are widely allowed. The barrier wall surface 400 preferably has a shape in which the inner peripheral edge B2 and the boundary B3 are joined from a transverse cross-sectional view, from the viewpoint of more reliably preventing the cultured cells from being present in the outer peripheral portion of the inner peripheral edge B2 when viewed from the upper and lower directions. A straight line or a shape that exists above the line.

Hereinafter, the second embodiment to the seventh embodiment in which the shape of the barrier surface 400 is different from that of the first embodiment will be described with reference to Figs. 10 to 15 . 10 to 15 correspond to Fig. 4 of the first embodiment. In the following embodiments, differences from the first embodiment will be mainly described, and the description of the same points will be omitted.

[Second Embodiment]

Fig. 10 is an enlarged view of a main part of a culture container according to a second embodiment. The barrier surface 400f shown in FIG. 10 is composed of a lower barrier surface portion 410f and an upper barrier surface portion 420f. The lower barrier surface portion 410f is raised from the base edge B2 at an angle θ 1 (90 degrees) to the bottom surface 200, and is connected to the upper barrier faucet portion 420f in parallel with the bottom surface 200 (the angle θ 2 is 180 degrees). Further, in FIG. 10, the ridge line is formed at the boundary between the lower barrier surface portion 410f and the upper barrier surface portion 420f, but the boundary may be subjected to C chamfering or R chamfering.

[Third embodiment]

Fig. 11 is an enlarged view of a main part of a culture container according to a third embodiment. The barrier surface 400g shown in FIG. 11 is composed of a lower barrier surface portion 410g and an upper barrier surface portion 420g. The lower barrier surface portion 410g raises the inner peripheral edge B2 as a base end at an angle θ 1 to the bottom surface 200, and the bottom and bottom The upper surface barrier faces 420g are connected in parallel with each other (the angle θ 2 is 180 degrees).

In the present embodiment, the angle θ 1 may exceed 90 degrees and be 150 degrees or less. When the upper limit is exceeded, the entire region in the inner periphery B2 to be cultured and observed can be observed with good visibility, and the culture cells can be reduced by "upper and lower limits". It is difficult to observe the outer portion of the inner circumference B2 when viewed in the direction". Further, in FIG. 11, the ridge line is formed at the boundary between the lower barrier surface portion 410g and the upper barrier surface portion 420g, but the boundary may be subjected to C chamfering or R chamfering.

[Fourth embodiment]

Fig. 12 is an enlarged view of a main part of a culture container according to a fourth embodiment. The barrier surface 400h shown in FIG. 12 is composed of a lower barrier surface portion 410h and an upper barrier portion 420h. The angle θ 1 between the lower barrier surface portion 410h and the bottom surface 200 is the same as the angle θ 2 formed by the upper barrier surface portion 420h and the bottom surface 200. Therefore, the two surfaces are connected without borders. Therefore, the structure of the barrier surface 400h is simple, and it is preferable from the viewpoint of manufacturing.

[Fifth Embodiment]

Fig. 13 is an enlarged view of a main part of a culture container according to a fifth embodiment. The cross-sectional shape of the barrier surface 400i shown in Fig. 13 from the lateral direction is a curved surface. The angles θ, θ', and θ" formed by the tangent of the curved surface and the bottom surface 200 gradually increase from the side of the bottom surface 200 toward the side of the peripheral side wall surface 300 (θ < θ' < θ"). Thereby, it is possible to more reliably prevent the cultured cells from being present in the "outer side portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up-and-down direction", and the structure of the barrier rib surface 400i itself is also simple, and it is preferable from the viewpoint of manufacture. Further, the angles θ, θ', and θ" may be changed in a range of 90 degrees or more and 180 degrees or less.

[Sixth embodiment]

Fig. 14 is an enlarged view of a main part of the culture container according to the sixth embodiment. The barrier surface 400j shown in Fig. 14 is composed of a lower barrier surface portion 410j and an upper barrier portion 420j. The cross-sectional shape of the lower barrier surface portion 410j viewed from the lateral direction is a straight line, and the inner peripheral edge B2 is raised as a base end so as to be at an angle θ1 from the bottom surface 200, and the position at the height h1 and the cross-sectional shape are curved upper side barrier faces. 420j connection settings. At a position of the height h1, a ridge line is formed at a boundary between the lower barrier rib portion 410j and the upper barrier façade 420j.

In the present embodiment, the angle θ 1 may be 90 degrees or more and 150 degrees or less. When viewed from the upper and lower sides, the entire area in the inner peripheral edge B2 to be cultured and observed can be observed with good visibility, and can be more reliably made lower than the upper limit value. The cultured cells are not present in the "outer side portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up-and-down direction". The cross-sectional shape of the upper barrier surface portion 420j may be the same curved surface as the barrier surface 400i of the fifth embodiment.

[Seventh embodiment]

Fig. 15 is an enlarged view of a main part of a culture container according to a seventh embodiment. The barrier surface 400k shown in Fig. 15 is composed of a lower barrier surface portion 410k and an upper barrier surface portion 420k. The lower barrier surface portion 410k is raised with the inner peripheral edge B2 as a base end so as to be at an angle θ1 from the bottom surface 200, and is connected to the upper barrier façade 420k at a position of the height h1. At the position of the height h1, the ridge line is formed at the boundary between the lower barrier surface portion 410k and the upper barrier surface portion 420k. The upper barrier surface portion 420k is inclined toward the peripheral side wall surface 300 at an angle θ 2 with respect to the bottom surface 200, and forms a boundary B3 with the circumferential side wall surface 300.

In the present embodiment, the angle θ 1 may be 90 degrees or more and 150 degrees or less. When viewed from the upper and lower directions by being at least the lower limit value, the visual field can be observed well and the culture should be observed. The entire region in the inner peripheral edge B2 of the observation is such that the cultured cells are reduced to the "outer portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up-and-down direction" by being equal to or less than the upper limit value.

The angle θ 2 is not particularly limited. For example, there is a tendency that the larger the angle θ 2 is, the more the height h1 of the lower barrier quotient portion 410k can be sufficiently ensured, and the cultured cells are reduced to the "outer side portion of the inner peripheral edge B2 which is difficult to observe when viewed in the up and down direction"; the angle θ 2 The smaller the size, the more the volume of the culture solution to be contained can be sufficiently obtained; the operator can appropriately determine the two tendencies based on these two tendencies.

In addition, in FIG. 15, the ridge line is formed in the boundary between the lower side barrier surface portion 410k and the upper side barrier surface portion 420k, but the boundary may be subjected to C chamfering or R chamfering.

<Modification>

Further, in the second to seventh embodiments described above, modifications are possible similarly to the first embodiment. In other words, as long as at least a part of the meniscus M (see FIGS. 3 to 9) is provided outside the inner periphery B2, the boundary E (see FIGS. 3 to 9) is not formed on the barrier faces 400f and 400g. 400h, 400i, 400j, 400k, the lower side barrier surfaces 410f, 410j, which are the base end B2 as the base end and rise at 90 degrees with respect to the bottom surface 200, may be allowed to be arbitrary structure. In other words, in the modification, the liquid surface S (see FIGS. 3 to 9) of the culture solution may be configured when the culture solution is stored in the culture chamber C (see FIGS. 3 to 9). The boundary E of the inner wall of the culture vessel becomes outside the inner peripheral edge B2.

[Eighth Embodiment]

Fig. 16 is a schematic perspective view showing the culture container of the eighth embodiment. The culture container 1001 has a plurality of culture chambers C. In the present embodiment, one culture container 1001 has six culture chambers C, but the number of culture chambers C is not limited thereto. For example, there is a tendency that the number of culture chambers C is higher. In many cases, it is preferable to enjoy the effect of concentrating a large part of the cultured cells on the observable portion; by limiting the amount of the culture chamber C to some extent, the bottom surface for cultivation and observation can be appropriately obtained. In the space above 200, the operator can appropriately determine the number of the culture chamber C based on the above situation. The structure of the culture chamber C of the culture container 1001 can be applied to any of the first to seventh embodiments and the modifications thereof.

[Examples]

The invention is specifically illustrated by the following examples, but the invention is not limited to the following examples.

<Example 1>

The structure shown in FIG. 9 was produced by laminating a circular plate of polystyrene to a cut product using polycarbonate (cutting on the inner peripheral side of the bottomless cylinder without a cover). The test piece of the culture container is further produced (in addition, the official product of the culture container can also be produced by integral molding. Hereinafter, the test piece produced by lamination is also simply referred to as "culture container"). θ 1 is 90 degrees and θ 2 is 120 degrees.

3 mL of a medium containing 10% serum was added to the culture chamber of the prepared culture vessel, and the culture chamber was observed from the up and down direction. Fig. 17 shows a photograph viewed in the up and down direction. As shown in Fig. 17, the culture container has the barrier surface 400e, and therefore most of the meniscus M is generated on the outer side of the inner periphery B2a, whereby it can be confirmed that the inside of the inner periphery B2a for cultivation and observation is well recognized. The range was expanded as compared with Comparative Example 1 below.

3 mL of medium containing 10% serum and 384,000 human fibroblasts were added to the culture chamber of the prepared culture vessel, and then cultured at 37 ° C under 5% CO ₂ for 24 hours, and observed under a microscope with a magnification of 40 times. The central part of the room and the surrounding area. Fig. 18 is a microscopic observation photograph of the central portion of the culture chamber, and Fig. 19 is a microscopic observation photograph of the peripheral portion of the culture chamber. As shown in Fig. 18, it was confirmed that the cells were normally followed by the bottom surface 200. Further, as shown in Fig. 19, it can be confirmed that most of the meniscus M is generated on the outer side of the inner peripheral edge B2a due to the barrier surface 400e, and the inner observable range for the inner periphery B2a for culture and observation is compared with the following. Example 1 is expanded.

<Example 2>

The structure shown in FIG. 9 was produced by laminating a circular plate of polystyrene to a cut product using polycarbonate (cutting on the inner peripheral side of the bottomless cylinder without a cover). Cultivate the test piece of the container. The culture container of this example was the same as the culture container of Example 1 except that θ 2 was 150 degrees.

3 mL of a medium containing 10% serum was added to the culture chamber of the prepared culture vessel, and the culture chamber was observed from the up and down direction. Fig. 20 shows a photograph viewed in the up and down direction. As shown in Fig. 20, it was confirmed that the culture container has the barrier surface 400e, and the shadow formed by the meniscus M is generated on the outer side of the inner periphery B2a, and can be clearly recognized inside the inner periphery B2a for cultivation and observation. The range was expanded as compared with Comparative Example 1 below.

3 mL of medium containing 10% serum and 384,000 human fibroblasts were added to the culture chamber of the prepared culture vessel, and then cultured at 37 ° C under 5% CO ₂ for 24 hours, and observed under a microscope with a magnification of 40 times. The central part of the room and the surrounding area. Fig. 21 is a photomicrograph of the central portion of the culture chamber, and Fig. 22 is a photomicrograph of the peripheral portion of the culture chamber. As shown in Fig. 21, it was confirmed that the cells were normally followed by the bottom surface 200. Further, as shown in Fig. 22, it can be confirmed that most of the meniscus M is generated on the outer side of the inner peripheral edge B2a due to the barrier surface 400e, and the range which can be observed inside the inner periphery B2a for cultivation and observation is compared with the following. Example 1 is expanded.

Microscopic observation of the peripheral portion of the culture chamber of Example 1 and Example 2 Comparing the sheets (Figs. 19 and 22), it was confirmed that the range of cells that can be clearly visualized in Example 2 (Fig. 22) was wider. In the second embodiment, the boundary Ee between the culture liquid surface S and the inner wall surface of the culture container 100e is located on the upper peripheral side wall surface portion 420e, and further, the specific height hd of the barrier surface 400e is lower than the culture liquid contained in the culture chamber C. The water level H is also below the height of the boundary E. Therefore, the range of the meniscus M generated on the outer side of the inner periphery B2a in the second embodiment is larger as compared with the first embodiment in which θ 2 is 120 degrees, and the present embodiment 2 ( In Fig. 22), the range in which cells can be clearly recognized is made wider.

<Comparative Example 1>

A culture vessel having no barrier surface was produced by laminating a circular plate of polystyrene to a lidless bottomless cylinder of polycarbonate. A schematic cross-sectional view of the culture vessel is shown in FIG.

3 mL of a medium containing 10% serum was added to the culture chamber of the prepared culture vessel, and the culture chamber was observed from the up and down direction. Fig. 24 shows a photograph viewed in the up and down direction. As shown in Fig. 24, the meniscus M was all generated inside the peripheral edge B, and the shadow formed by the meniscus M was confirmed inside the periphery B for cultivation and observation.

3 mL of medium containing 10% serum and 384,000 human fibroblasts were added to the culture chamber of the prepared culture vessel, and then cultured at 37 ° C under 5% CO ₂ for 24 hours, and observed under a microscope with a magnification of 40 times. The central part of the room and the surrounding area. Fig. 25 is a photomicrograph of the central portion of the culture chamber, and Fig. 26 is a photomicrograph of the peripheral portion of the culture chamber. As shown in Fig. 25, it was confirmed that the cells were normally followed by the bottom surface 200. Further, as shown in Fig. 26, it was confirmed that all of the meniscus M was generated inside the peripheral edge B, and a shadow formed by the meniscus M was generated inside the periphery B for culture and observation, and it was difficult to observe cells near the periphery B. The range that can be observed inside the periphery B used for culture and observation is narrow.

[Correspondence between each component in the first embodiment to the eighth embodiment and each component of the request item]

In the present invention, the culture containers 100 and 1001 correspond to a "culture container", the bottom surface 200 corresponds to the "bottom surface", and the peripheral side wall surfaces 300, 300b, and 300d correspond to the "peripheral side wall surface", and the barrier surfaces 400, 400a, 400b, and 400c 400d, 400e, 400f, 400g, 400h, 400i, 400j, and 400k correspond to the "barrier surface", and the lower barrier surface portions 410, 410f, 410g, 410h, 410j, and 410k correspond to the "lower barrier surface portion" and the upper barrier surface portion. 420, 420d, 420e, 420f, 420g, 420h, 420j, and 420k correspond to "upper barrier face", peripheral edge O corresponds to "perimeter", inner periphery B2, B2a corresponds to "internal circumference", and boundaries B3, B3b, and B3d are equivalent. In the "boundary" of the (barrier surface and the peripheral side wall surface), the culture chamber C corresponds to the "culture chamber", and the angle θ 1 corresponds to the "angle" (the angle formed by the lower barrier surface portion and the bottom surface), the angle θ, θ', θ" corresponds to (the angle formed by the tangent to the bottom surface of the cross-sectional shape of the barrier surface).

The preferred embodiments of the present invention are as described above, but the present invention is not limited to the embodiments, and various other embodiments are possible without departing from the spirit and scope of the invention. Furthermore, the actions and effects described in the present embodiment are merely examples and are not intended to limit the present invention.

[Industrial availability]

According to the present invention, it is possible to provide a culture container in which the shadow formed by the meniscus does not interfere with the observation of the cultured cells even if a meniscus is generated.

100‧‧‧ culture container

200‧‧‧ bottom

300‧‧‧ Weekly side wall

400‧‧ ‧ barrier wall

B2‧‧‧ (bottom) internal circumference

B3‧‧‧ (Bound wall and circumferential side wall) boundary

C‧‧‧Cultivation room

Claims (6)

A culture container comprising a culture chamber having a bottom surface and a circumferential side wall surface provided on a periphery of the bottom surface, wherein a barrier wall surface of a specific height is disposed around the circumference in the culture chamber, and the barrier surface is Formed as follows: an inner peripheral edge on the bottom surface that is further inside than the peripheral edge is raised as a base end, and is formed to face the peripheral side wall surface and form a boundary with the peripheral side wall surface. The culture container according to claim 1, wherein the shape of the barrier surface is a straight line, a curved line, and a combination of a straight line and a curved line when viewed from a transverse cross section. The culture container according to claim 1 or 2, wherein, in a transverse cross-sectional view, the barrier wall surface is provided by a lower barrier surface portion rising from the inner peripheral edge and connected to the lower barrier surface portion and formed on the circumferential side The side of the wall surface is formed by the upper barrier surface portion, and the side barrier surface portion is inclined at an angle of 90 degrees or more and 150 degrees or less from the bottom surface from the inner peripheral edge. The culture container according to claim 3, wherein the lower side barrier surface portion having the same slope from each other is connected to the upper barrier surface portion without a boundary from a transverse cross-sectional view. The culture container according to the second aspect of the invention, wherein the shape of the barrier wall surface in the cross-sectional view is a curve, and an angle formed by a tangent line of the curve and the bottom surface gradually increases from the bottom surface side toward the circumferential side wall surface side. . The culture container according to any one of claims 1 to 5, wherein the circumferential side wall surface and the inner peripheral edge have a circular shape in plan view, and the inner peripheral edge has a diameter of 30 mm or more and 170 mm or less.