WO2016047751A1 - Récipient de culture - Google Patents

Récipient de culture Download PDF

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Publication number
WO2016047751A1
WO2016047751A1 PCT/JP2015/077086 JP2015077086W WO2016047751A1 WO 2016047751 A1 WO2016047751 A1 WO 2016047751A1 JP 2015077086 W JP2015077086 W JP 2015077086W WO 2016047751 A1 WO2016047751 A1 WO 2016047751A1
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WIPO (PCT)
Prior art keywords
culture
peripheral edge
barrier surface
inner peripheral
barrier
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PCT/JP2015/077086
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English (en)
Japanese (ja)
Inventor
林　大輔
春男 大久保
魚住　孝之
宏昭 紀伊
Original Assignee
住友ベークライト株式会社
株式会社ニコン
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Application filed by 住友ベークライト株式会社, 株式会社ニコン filed Critical 住友ベークライト株式会社
Publication of WO2016047751A1 publication Critical patent/WO2016047751A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/22—Petri dishes

Definitions

  • the present invention relates to a culture vessel. More specifically, the present invention relates to a culture vessel having a meniscus effect.
  • This application claims priority based on Japanese Patent Application No. 2014-195164 filed in Japan on September 25, 2014, the contents of which are incorporated herein by reference.
  • Patent Document 1 discloses that a solution that becomes a gel state by heating containing a biological substance on the bottom surface of a culture vessel is not yet available. Disclosed is a cell culture device having a layer A frozen in a gelled state, and a layer B frozen on an aqueous solution that does not become a gel state by heating. Has been.
  • the conventional technique aimed at solving the meniscus problem is performed by means for producing an action for reducing or preventing meniscus generation itself.
  • an object of the present invention is to provide a culture vessel that allows generation of meniscus and can solve the problem of meniscus.
  • the inventor has found that the object of the present invention can be achieved by providing the culture vessel with a structure that allows the meniscus to escape outside the range to be cultured and observed, and to complete the present invention. It came.
  • the culture container of the present invention includes a culture chamber having a bottom surface and a peripheral side wall surface provided on the periphery of the bottom surface.
  • a barrier surface is provided along the periphery of the bottom surface. The barrier surface rises with the inner peripheral edge inside the peripheral edge on the bottom surface as a base end, and is formed so as to go to the peripheral side wall surface and to form a boundary with the peripheral side wall surface.
  • the barrier surface is set to a height that allows a drop to be formed to a degree that suppresses cell survival on the barrier surface.
  • the shape of the barrier surface in a cross-sectional view from the lateral direction may be any of a straight line, a curved line, and a combination of a straight line and a curved line.
  • the barrier surface is composed of 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 side wall surface. It is preferable that the lower barrier surface portion rising from the angle forms an angle of 90 degrees or more and 150 degrees or less with the bottom surface.
  • the angle of the lower barrier surface portion is 90 degrees or more, the observation range of the collected cultured cells can be expanded without obstructing the field of view except for the peripheral portion in the culture chamber, which is 150 degrees or less. Thus, it is possible to further reduce the invasion of the cultured cells to the peripheral portion in the culture chamber that is difficult to observe.
  • the lower barrier surface portion and the upper barrier surface portion having the same inclination may be continuously provided without a boundary in a cross-sectional view from the lateral direction.
  • the structure of the barrier surface is simple, which is preferable from the viewpoint of manufacturing.
  • the curve has a shape in which an angle formed between the tangent and the bottom surface increases from the bottom surface side toward the peripheral side wall surface side. That is, the shape of the barrier surface in a cross-sectional view from the lateral direction (direction parallel to the bottom surface) is a curve, and the curve is an angle formed between the tangent line of the curve and the bottom surface from the bottom surface side toward the peripheral side wall surface side.
  • the shape may gradually increase.
  • a top view shape of the peripheral side wall surface and the inner peripheral edge may be a circle, and in this case, the diameter of the inner peripheral edge may be not less than 30 mm and not more than 170 mm.
  • the diameter of the inner peripheral edge is 30 mm or more, a space on the bottom surface for culturing and observation can be preferably obtained, and when it is 170 mm or less, an effect of collecting most of the cultured cells in an observable part. Can be enjoyed preferably.
  • a culture container in which a shadow formed by the meniscus does not hinder the observation of cultured cells can be provided.
  • FIG. 3 shows a partially enlarged view of the schematic cross-sectional view of FIG. 2.
  • the principal part enlarged view of the typical sectional drawing of FIG. 2 is shown.
  • the typical sectional view at the time of use of the culture container of the 1st modification of a 1st embodiment is shown.
  • the typical sectional view at the time of use of the culture container of the 2nd modification of a 1st embodiment is shown.
  • the typical sectional view at the time of use of the culture container of the 3rd modification of a 1st embodiment is shown.
  • the typical sectional view at the time of use of the culture container of the 4th modification of a 1st embodiment is shown.
  • the typical sectional view at the time of use of the culture container of the 5th modification of a 1st embodiment is shown.
  • the principal part enlarged view of the culture container of 2nd Embodiment is shown.
  • the principal part enlarged view of the culture container of 3rd Embodiment is shown.
  • the principal part enlarged view of the culture container of 4th Embodiment is shown.
  • the principal part enlarged view of the culture container of 5th Embodiment is shown.
  • the principal part enlarged view of the culture container of 6th Embodiment is shown.
  • the principal part enlarged view of the culture container of 7th Embodiment is shown.
  • FIG. 2 is a vertical view photograph after adding a culture medium to the culture container prepared in Example 1.
  • FIG. 2 is a vertically viewed photograph (partially enlarged) after cell culture in the culture vessel prepared in Example 1.
  • FIG. 2 is a vertically viewed photograph (partially enlarged) after cell culture in the culture vessel prepared in Example 1.
  • FIG. 4 is a vertical view photograph after adding a culture medium to the culture container produced in Example 2.
  • FIG. 4 is a vertically viewed photograph (partially enlarged) after cell culture in a culture vessel prepared in Example 2.
  • FIG. 4 is a vertically viewed photograph (partially enlarged) after cell culture in a culture vessel prepared in Example 2.
  • FIG. 4 is a vertically viewed photograph (partially enlarged) after cell culture in a culture vessel prepared in Example 2.
  • the typical sectional view of the culture container produced by comparative example 1 is shown. It is an up-down direction photograph after adding a culture medium to the culture container produced by the comparative example 1. It is an up-down direction view photograph (partially enlarged) after cell culture in the culture container produced in Comparative Example 1. It is an up-down direction view photograph (partially enlarged) after cell culture in the culture container produced in Comparative Example 1.
  • FIG. 1 the typical external appearance perspective view of the culture container of 1st Embodiment is shown.
  • FIG. 2 the typical sectional drawing at the time of use of the culture container of 1st Embodiment is shown.
  • the schematic cross-sectional view of FIG. 2 is a view of a plane cut along a vertical plane including the line AA of FIG.
  • FIG. 3 shows a partial enlargement of the schematic cross-sectional view of FIG.
  • FIG. 4 shows an enlarged view of a main part (circled portion) of the schematic cross-sectional view of FIG.
  • the upper side of each drawing is referred to as the upper side
  • the lower side is referred to as the lower side.
  • the culture vessel 100 includes a bottom surface 200, a peripheral wall surface 300, and a barrier surface 400 as surfaces constituting the inside thereof.
  • the culture chamber C is defined by the bottom surface 200, the peripheral side wall surface 300 and the barrier surface 400.
  • the culture container of the present embodiment includes a bottom surface 200, a barrier surface 400 provided outside along the peripheral edge B2 of the bottom surface 200, and a peripheral side wall surface provided further outward along the peripheral edge of the barrier surface 400. 300 culture chamber C.
  • the bottom surface 200 has a circular shape when viewed in the vertical direction, and is provided with a peripheral side wall surface 300 that rises from the entire periphery of the peripheral edge O and opens upward as shown in FIG.
  • the barrier surface 400 provided around the entire periphery of the peripheral edge O of the bottom surface 200 at a predetermined height h rises from the peripheral edge O of the bottom surface 200 to the inside with a predetermined distance w as a base end, While going to the peripheral side wall surface 300, it is formed so as to form a boundary B3 with the peripheral side wall surface 300 at a predetermined height h.
  • the predetermined distance w corresponds to the distance between the peripheral edge O and the internal peripheral edge B2.
  • the predetermined height h corresponds to the distance between the peripheral edge O and the boundary B3, and is set so as to be able to configure a shape that suppresses cell remaining on the barrier surface 400 in relation to the predetermined distance w.
  • the temperature is set to be equal to or lower than the lowest water level H of the culture solution to be stored in the culture chamber C.
  • the field of view from the vertical direction is difficult to observe because the culture liquid surface S in the peripheral part of the culture chamber C is curved due to the meniscus M phenomenon.
  • the culture vessel 100 by providing the barrier surface 400 having a predetermined height h rising from the inner peripheral edge B2, the culture cell is prevented from moving from the inner peripheral edge B2 to the barrier surface 400, and the barrier surface 400 is moved from the inner peripheral edge B2.
  • the barrier surface 400 By separating the inner peripheral edge B2 and the peripheral side wall surface 300 by a predetermined distance w in the outward direction, at least a part of the portion where the meniscus M is generated escapes from the inner peripheral edge B2. Therefore, while allowing the generation of the meniscus M, most of the cultured cells can be present inside the observable inner peripheral edge B2.
  • the predetermined distance w is not particularly limited.
  • the larger the predetermined distance w the more the influence of the meniscus M tends to be reduced.
  • the smaller the predetermined distance w the more the space on the bottom surface 200 for culture and observation tends to be sufficient.
  • the predetermined distance w may be 2 mm or more and 3 mm or less.
  • the predetermined distance w is set to be equal to or larger than the width W of the meniscus M.
  • the predetermined height h is not particularly limited, as long as it can be set so as to be able to configure a drop that suppresses cell remaining on the barrier surface 400 in relation to the predetermined distance w.
  • the predetermined height h is 1 mm or more and 21 mm or less, preferably 1 mm or more and 15 mm or less.
  • the predetermined height h is set to be equal to or lower than the water level H of the culture solution to be stored in the culture chamber C from the viewpoint of taking a sufficient volume to store the culture solution.
  • the boundary E between the culture liquid surface S and the inner wall surface of the culture vessel 100 is on the peripheral wall surface 300.
  • the barrier surface 400 includes a lower barrier surface portion 410 and an upper barrier surface portion 420.
  • the lower barrier surface portion 410 rises with the inner peripheral edge B2 as a base end so as to form an angle ⁇ 1 with the bottom surface 200, and is connected to the upper barrier surface portion 420 at a height h1.
  • the boundary between the lower barrier surface portion 410 and the upper barrier surface portion 420 forms a ridge line.
  • the upper barrier surface portion 420 is inclined toward the peripheral side wall surface 300 at an angle ⁇ 2 with respect to the bottom surface 200, and forms a boundary B3 with the peripheral side wall surface 300.
  • the angle ⁇ 1 between the lower barrier surface portion 410 and the bottom surface 200 is 90 degrees. Thereby, it is possible to effectively prevent the cultured cells on the bottom surface 200 from moving beyond the inner peripheral edge B2.
  • the angle ⁇ 2 that is the inclination angle of the upper barrier surface portion 420 is not particularly limited. For example, as the angle ⁇ 2 is larger, the height h1 of the lower barrier surface portion 410 is sufficiently secured, and the invasion of the cultured cells into the outer portion of the inner peripheral edge B2 that is difficult to observe in the vertical direction can be reduced. It can be determined appropriately by those skilled in the art based on the tendency that the smaller the angle ⁇ 2 is, the more the volume for storing the culture solution can be taken.
  • the height h1 of the lower barrier surface portion 410 is not particularly limited.
  • the height h1 may be set to, for example, 0.5 mm or 1 mm.
  • the diameter ⁇ of the inner peripheral edge B2 may be equal to the diameter of the inner bottom surface of the conventional culture vessel. 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 above the lower limit value, it is possible to preferably take a space on the bottom surface 200 for culture and observation, and by being below the upper limit value, an effect of collecting most of the cultured cells in an observable portion. Can be enjoyed preferably.
  • the culture vessel 100 can be made of a transparent material.
  • the material having transparency include inorganic substances typified by glass and quartz, and organic substances typified by synthetic resins.
  • Synthetic resins include polystyrene (PS), polypropylene (PP), polymethylpentene (PMP), polycarbonate (PC), polymethyl methacrylate (PMMA), polymethylacrylmethylimide (PMMI), and cycloolefin copolymer (COC). And the like.
  • combined from two or more of the monomer units of these polymers is also mentioned.
  • the culture vessel 100 may be a structure formed by integral molding, or may be a structure formed by a combination of a member forming the bottom surface 200 and members forming the peripheral wall surface 300 and the barrier surface 400, for example.
  • each member may be made of different materials.
  • the bottom surface 200, the peripheral wall surface 300, and the barrier surface 400 of the culture vessel 100 may be subjected to a surface treatment from the viewpoint of physical, chemical, and / or biochemistry.
  • a surface treatment can be appropriately selected by those skilled in the art.
  • the barrier surface 400 or the barrier surface 400 and the peripheral wall surface 300 may be subjected to a surface treatment for inhibiting cell culture.
  • an additional structure may be provided depending on the purpose of use of the culture vessel 100 or the like.
  • Such an additional structure can be appropriately selected by those skilled in the art.
  • the bottom surface 200 such as a lid for placing the culture chamber C in a covered state, a partition for partitioning the contents, and a grid pattern for specifying the amount and position of the contents (particularly cell groups).
  • the guide structure provided in the above.
  • the culture vessel 100 can be variously modified in addition to the illustrated one.
  • at least a part of the meniscus M only needs to escape to the outside of the inner peripheral edge B2, so that the boundary E is 90 degrees with respect to the bottom surface 200 with the inner peripheral edge B2 being the base end of the barrier surface 400.
  • Any structure is acceptable as long as it is not formed on the rising lower barrier surface 410. That is, in the modification, when the culture solution is accommodated in the culture chamber C, the boundary E between the liquid surface S of the culture solution and the inner wall of the culture vessel 100 is outside the inner peripheral edge B2. Good.
  • FIG. 5 shows a first modification of the first embodiment.
  • the barrier surface 400a of the culture vessel 100a shown in FIG. 5 is set such that the predetermined distance wa is shorter than the predetermined distance w in the first embodiment, so that the inner peripheral edge B2a serving as the base end is the inner space in the first embodiment. It is outside the peripheral edge B2.
  • the meniscus M that escapes to the outside from the inner peripheral edge B2a becomes a part thereof, but the influence of the meniscus M can be reduced. That is, also in this modified example, the meniscus M is escaped to the outside from the inner peripheral edge B2a, so that the influence of the meniscus M can be reduced.
  • FIG. 6 shows a second modification of the first embodiment.
  • the barrier surface 400b of the culture vessel 100b shown in FIG. 6 has a predetermined height hb set to be larger than the predetermined height h in the first embodiment, so that the boundary B3b with the peripheral wall surface 300b is the first implementation. It is above the boundary B3 in the form. As a result, it is possible to further reduce the invasion of the cultured cells into the outer portion of the inner peripheral edge B2, which is difficult to observe when viewed in the vertical direction.
  • the predetermined 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 less than the height of the boundary E between the culture solution surface S and the peripheral side wall surface 300b.
  • FIG. 7 shows a third modification of the first embodiment.
  • the barrier surface 400c of the culture vessel 100c shown in FIG. 7 has an inner peripheral edge serving as a base end when the predetermined distance wa is set shorter than the predetermined distance w in the first embodiment.
  • B2a is outside the inner peripheral edge B2 in the first embodiment.
  • the barrier surface 400c has a boundary B3b with the peripheral side wall surface 300b above the boundary B3 in the first embodiment, as in the second modification.
  • the predetermined 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 less than the height of the boundary E between the culture solution surface S and the peripheral side wall surface 300b.
  • FIG. 8 shows a fourth modification of the first embodiment.
  • the barrier surface 400d of the culture vessel 100d shown in FIG. 8 is set such that the predetermined height hd is larger than the predetermined height hb in the second modified example, so that the boundary B3d with the peripheral wall surface 300d is the second. It is further above the boundary B3b in the modification. As a result, it is possible to further reduce the invasion of the cultured cells into the outer portion of the inner peripheral edge B2, which is difficult to observe when viewed in the vertical direction.
  • the predetermined height hd of the barrier surface 400d is to be accommodated in the culture chamber C.
  • the liquid level exceeds H and exceeds the height of the boundary Ed.
  • the meniscus M can escape to the outside of the inner peripheral edge B2, and the influence of the meniscus M can be avoided.
  • FIG. 9 shows a fifth modification of the first embodiment.
  • the barrier surface 400e of the culture vessel 100e shown in FIG. 9 is set such that the predetermined height hd is larger than the predetermined height hb in the second and third modifications.
  • the boundary B3d with the peripheral side wall surface 300d is further above the boundary B3b in the second and third modified examples.
  • the predetermined height hd of the barrier surface 400e is to be accommodated in the culture chamber C.
  • the liquid level exceeds H and exceeds the boundary Ee.
  • the predetermined distance wa is set shorter than the predetermined distance w in the first embodiment, so that the inner peripheral edge B2a serving as the base end is more than the inner peripheral edge B2 in the first embodiment.
  • the meniscus M that escapes to the outside from the inner peripheral edge B2a becomes a part thereof, but the influence of the meniscus M can be reduced. That is, also in this modified example, the meniscus M is escaped to the outside from the inner peripheral edge B2a, so that the influence of the meniscus M can be reduced.
  • the barrier surface 400 only needs to have a shape that passes between the inner peripheral edge B2 and the boundary B3 (a shape that connects the inner peripheral edge B2 and the boundary B3) in a cross-sectional view from the lateral direction. It is not limited to the shape shown in one embodiment.
  • the boundary between the lower barrier surface portion 410 and the upper barrier surface portion 420 may form a ridge line as shown in FIG. 4, or the boundary may be C chamfered or R chamfered.
  • the cross-sectional shape viewed from the lateral direction of the barrier surface 400 may be a straight line, a curved line, or a combination of a straight line and a curved line, and various shapes are widely acceptable. Is done.
  • the barrier surface 400 has the inner peripheral edge B2 and the boundary B3 in the cross-sectional view from the lateral direction, from the viewpoint of more surely preventing the cultured cells from being present on the outer portion of the inner peripheral edge B2, which is difficult to observe in the vertical direction.
  • FIGS. 10 to 15 correspond to FIG. 4 of the first embodiment.
  • differences from the first embodiment will be mainly described, and description of the same points will be omitted.
  • FIG. 10 shows an enlarged view of the main part of the culture vessel of the second embodiment.
  • the barrier surface 400f shown in FIG. 10 includes a lower barrier surface portion 410f and an upper barrier surface portion 420f.
  • the lower barrier surface portion 410f rises with the inner peripheral edge B2 as a base end so as to form an angle ⁇ 1 (90 degrees) with the bottom surface 200, and is connected to the upper barrier surface portion 420f parallel to the bottom surface 200 (angle ⁇ 2 is 180 degrees).
  • angle ⁇ 1 90 degrees
  • the boundary between the lower barrier surface portion 410f and the upper barrier surface portion 420f forms a ridge line, but the boundary may be C-chamfered or R-chamfered.
  • FIG. 11 shows the principal part enlarged view of the culture container of 3rd Embodiment.
  • the barrier surface 400g shown in FIG. 11 includes a lower barrier surface portion 410g and an upper barrier surface portion 420g.
  • the lower barrier surface portion 410g rises with the inner peripheral edge B2 as a base end so as to form an angle ⁇ 1 with the bottom surface 200, and is connected to the upper barrier surface portion 420g parallel to the bottom surface 200 (angle ⁇ 2 is 180 degrees).
  • the angle ⁇ 1 may be greater than 90 degrees and less than or equal to 150 degrees.
  • the boundary between the lower barrier surface portion 410g and the upper barrier surface portion 420g forms a ridge line, but the boundary may be C-chamfered or R-chamfered.
  • FIG. 12 shows an enlarged view of the main part of the culture vessel of the fourth embodiment.
  • the barrier surface 400h shown in FIG. 12 includes a lower barrier surface portion 410h and an upper barrier surface portion 420h. Since the angle ⁇ 1 formed by the lower barrier surface portion 410h and the bottom surface 200 is the same as the angle ⁇ 2 formed by the upper barrier surface portion 420h and the bottom surface 200, both surfaces are connected without boundaries. For this reason, the structure of the barrier surface 400h is simple, which is preferable from the viewpoint of manufacturing.
  • FIG. 13 shows the principal part enlarged view of the culture container of 5th Embodiment.
  • the barrier surface 400i shown in FIG. 13 has a curved surface when viewed from the lateral direction.
  • the angles ⁇ , ⁇ ′, ⁇ ′′ formed by the tangent to the curved surface and the bottom surface 200 gradually increase from the bottom surface 200 side toward the peripheral side wall surface 300 ( ⁇ ⁇ ′ ⁇ ′′).
  • the angles ⁇ , ⁇ ′, and ⁇ ′′ can be changed in the range of 90 degrees to 180 degrees.
  • FIG. 14 shows the principal part enlarged view of the culture container of 6th Embodiment.
  • the barrier surface 400j shown in FIG. 14 includes a lower barrier surface portion 410j and an upper barrier surface portion 420j.
  • the lower barrier surface portion 410j has a straight cross-sectional shape viewed from the lateral direction, rises with the inner peripheral edge B2 as the base end so as to form an angle ⁇ 1 with the bottom surface 200, and the cross-sectional shape is a curve at a position of height h1. It is connected to a certain upper barrier surface portion 420j. At the position of the height h1, the boundary between the lower barrier surface portion 410j and the upper barrier surface portion 420j forms a ridge line.
  • the angle ⁇ 1 may be not less than 90 degrees and not more than 150 degrees.
  • the cross-sectional shape of the upper barrier surface portion 420j may be a curved surface similar to the barrier surface 400i in the fifth embodiment.
  • FIG. 15 shows the principal part enlarged view of the culture container of 7th Embodiment.
  • the barrier surface 400k shown in FIG. 15 includes a lower barrier surface portion 410k and an upper barrier surface portion 420k.
  • the lower barrier surface portion 410k rises with the inner peripheral edge B2 as a base end so as to form an angle ⁇ 1 with the bottom surface 200, and is continuous with the upper barrier surface portion 420k at a height h1.
  • the boundary between the lower barrier surface portion 410k and the upper barrier surface portion 420k forms a ridge line.
  • the upper barrier surface portion 420k is inclined toward the peripheral side wall surface 300 at an angle ⁇ 2 with respect to the bottom surface 200, and forms a boundary B3 with the peripheral side wall surface 300.
  • the angle ⁇ 1 may be not less than 90 degrees and not more than 150 degrees.
  • the entire region within the inner peripheral edge B2 to be cultured and observed can be observed with good visibility from the vertical direction view. It is possible to reduce the entry of cultured cells into the outer portion of the inner peripheral edge B2, which is difficult.
  • the angle ⁇ 2 is not particularly limited.
  • the angle ⁇ 2 is larger, the height h1 of the lower barrier surface portion 410k is sufficiently secured, and the tendency of the cultured cells to enter the outer portion of the inner peripheral edge B2, which is difficult to observe in the vertical direction, can be reduced. Therefore, the smaller the angle ⁇ 2, the more appropriate it can be determined by those skilled in the art based on the tendency that a sufficient volume for accommodating the culture solution tends to be taken.
  • the boundary between the lower barrier surface portion 410k and the upper barrier surface portion 420k forms a ridge line, but the boundary may be C-chamfered or R-chamfered.
  • FIG. 16 is a schematic external perspective view of the culture vessel of the eighth embodiment.
  • the culture vessel 100l has a plurality of culture chambers C.
  • one culture vessel 100l has six culture chambers C, but the number of culture chambers C is not limited to this.
  • the number of the culture chambers C is not limited to this.
  • Those skilled in the art can appropriately determine the space on the bottom surface 200 for observation based on the appropriate space.
  • the structure of the culture chamber C of the culture vessel 100l the structure in any of the first to seventh embodiments and their modifications may be applied.
  • Example 1 A prototype of a culture vessel having the structure shown in FIG. 9 was prepared by laminating a polystyrene disc to a cut product using polycarbonate (in which the inner peripheral surface side of an open and bottomless cylinder was cut). (Note that the actual product of the culture vessel may be produced by integral molding. Hereinafter, the prototype produced by bonding is also simply referred to as “culture vessel”). ⁇ 1 was 90 degrees and ⁇ 2 was 120 degrees.
  • FIG. 17 shows a vertical view photograph.
  • the culture container has a barrier surface 400e, so that most of the meniscus M is generated outside the inner peripheral edge B2a, so that the culture container is well visible inside the inner peripheral edge B2a for culture and observation. It was confirmed that the range which can be expanded from the comparative example 1 mentioned later.
  • FIG. 18 shows a microscopic observation photograph of the center of the culture chamber
  • FIG. 19 shows the periphery of the culture chamber.
  • the barrier surface 400e causes most of the meniscus M to be generated outside the inner peripheral edge B2a, and the range that can be observed inside the inner peripheral edge B2a for culture and observation is Comparative Example 1 described later. It was confirmed that it was expanding.
  • Example 2 A prototype of a culture vessel having the structure shown in FIG. 9 was prepared by laminating a polystyrene disc to a cut product using polycarbonate (in which the inner peripheral surface of an open-bottomed bottomless cylinder was cut). .
  • the culture container of this example is the same as the culture container of Example 1 except that ⁇ 2 is 150 degrees.
  • FIG. 20 shows a vertical view photograph.
  • the culture vessel has a barrier surface 400e, so that a shadow caused by the meniscus M is generated outside the inner peripheral edge B2a, and there is a range that can be favorably seen inside the inner peripheral edge B2a for culture and observation. It was confirmed that it was larger than Comparative Example 1 described later.
  • FIG. 21 shows a microscope observation photograph of the center of the culture chamber and FIG. 22 shows the periphery of the culture chamber. As shown in FIG. 21, it was confirmed that the cells normally adhered to the bottom surface 200. Further, as shown in FIG. 22, the barrier surface 400e causes most of the meniscus M to be generated outside the inner peripheral edge B2a, and the range that can be observed inside the inner peripheral edge B2a for culture and observation is Comparative Example 1 described later. It was confirmed that it was expanding.
  • Example 2 Comparison of the microscopic observation photographs (FIGS. 19 and 22) in the periphery of the culture chamber of Example 1 and Example 2 confirms that Example 2 (FIG. 22) has a wider range in which the cells can be seen well. It was done.
  • the boundary Ee between the culture liquid surface S and the inner wall surface of the culture vessel 100e is on the upper peripheral wall surface portion 420e, and the predetermined height hd of the barrier surface 400e is accommodated in the culture chamber C.
  • the level of the culture broth was below the level H and the height of the boundary E was also below.
  • Example 2 in which ⁇ 2 is 150 degrees, the range of meniscus M generated outside the inner peripheral edge B2a becomes larger, and this Example 2 ( In FIG. 22), the range in which the cells can be visually recognized is widened.
  • a culture vessel having no barrier surface was prepared by attaching a polystyrene disk to a polycarbonate open-bottomed bottomless cylinder. A schematic cross-sectional view of this culture vessel is shown in FIG.
  • FIG. 24 shows a vertical view photograph. As shown in FIG. 24, all of the meniscus M occurred inside the peripheral edge B, and a shadow due to the meniscus M was confirmed inside the peripheral edge B for culture and observation.
  • FIG. 25 shows a microscope observation photograph of the center of the culture chamber
  • FIG. 26 shows the periphery of the culture chamber. As shown in FIG. 25, it was confirmed that the cells normally adhered to the bottom surface 200.
  • FIG. 25 shows a microscope observation photograph of the center of the culture chamber
  • FIG. 26 shows the periphery of the culture chamber. As shown in FIG. 25, it was confirmed that the cells normally adhered to the bottom surface 200.
  • the culture vessels 100 and 100l correspond to “culture vessels”
  • the bottom surface 200 corresponds to “bottom surface”
  • the peripheral side wall surfaces 300, 300b, and 300d correspond to “peripheral side wall surfaces”
  • the barrier surface 400 400a, 400b, 400c, 400d, 400e, 400f, 400g, 400h, 400i, 400j, 400k correspond to the “barrier surface”
  • the lower barrier surface portions 410, 410f, 410g, 410h, 410j, 410k are “lower side”.
  • the upper barrier surface portions 420, 420d, 420e, 420f, 420g, 420h, 420j, 420k correspond to the “upper barrier surface portion”
  • the peripheral edge O corresponds to the “periphery”
  • the inner peripheral edges B2, B2a corresponds to the “inner periphery”
  • the boundaries B3, B3b, B3d correspond to “boundaries” (between the barrier surface and the peripheral side wall surface)
  • the culture chamber C is the “culture chamber”.
  • the angle ⁇ 1 corresponds to the “angle” (the lower barrier surface portion and the bottom surface form), and the angles ⁇ , ⁇ ′, and ⁇ ′′ (the tangent of the cross-sectional shape of the barrier surface and the bottom surface form) “ Corresponds to “Angle”.
  • a culture container in which a shadow formed by the meniscus does not hinder the observation of cultured cells can be provided.

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
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Abstract

L'invention concerne un récipient de culture qui contient une chambre de culture possédant une face fond, et une face paroi latérale périphérique agencée sur un bord périphérique de ladite face fond. Une face paroi barrière de hauteur prédéfinie, est agencée de manière périphérique suivant ledit bord périphérique, à l'intérieur de ladite chambre de culture. Ladite face paroi barrière se dresse avec pour extrémité de base un bord périphérique de partie interne côté interne par rapport audit bord périphérique, sur ladite face fond, et est formée de sorte que tout en se dirigeant vers ladite face paroi latérale périphérique, elle forme une limite par rapport à cette dernière.
PCT/JP2015/077086 2014-09-25 2015-09-25 Récipient de culture WO2016047751A1 (fr)

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JP2014195164 2014-09-25
JP2014-195164 2014-09-25

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WO2016047751A1 true WO2016047751A1 (fr) 2016-03-31

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017163378A1 (fr) * 2016-03-24 2017-09-28 株式会社ニコン Récipient de culture
CN112534038A (zh) * 2018-05-30 2021-03-19 太阳生物科学股份公司 一种用于培养生物材料的孔

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JPS58500972A (ja) * 1981-06-22 1983-06-23 バツクスター　トラベノル　ラボラトリーズ　インコーポレーテツド 生物医学分析用改良トレイ
JPS6251977A (ja) * 1985-09-02 1987-03-06 Terumo Corp 組織培養プレ−ト
WO1991006624A1 (fr) * 1989-10-26 1991-05-16 Costar Corporation Cuvette de fecondation in vitro
JPH05181068A (ja) * 1991-08-20 1993-07-23 Hoya Corp メニスカス制御用平板
JP2006280298A (ja) * 2005-04-01 2006-10-19 Nipro Corp 細胞培養用容器
JP2007504816A (ja) * 2003-09-10 2007-03-08 オバチュアー リサーチ インコーポレイティド 細胞、胚または、卵母細胞を取り扱う装置と方法
JP2008045979A (ja) * 2006-08-15 2008-02-28 Yokogawa Electric Corp マイクロプレート
JP2008532501A (ja) * 2005-03-10 2008-08-21 アレキサンダー，ロバート ウイルスの診断法およびこれに使用するウェル
JP2010527582A (ja) * 2007-05-30 2010-08-19 株式会社ニコン 培養容器
JP2010200748A (ja) * 2009-02-09 2010-09-16 Dainippon Printing Co Ltd 細胞培養容器
JP2015047136A (ja) * 2013-09-03 2015-03-16 大日本印刷株式会社 細胞培養容器

Patent Citations (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4321330A (en) * 1980-04-04 1982-03-23 Baker Fraser L Tissue culture device
JPS58500972A (ja) * 1981-06-22 1983-06-23 バツクスター　トラベノル　ラボラトリーズ　インコーポレーテツド 生物医学分析用改良トレイ
JPS6251977A (ja) * 1985-09-02 1987-03-06 Terumo Corp 組織培養プレ−ト
WO1991006624A1 (fr) * 1989-10-26 1991-05-16 Costar Corporation Cuvette de fecondation in vitro
JPH05181068A (ja) * 1991-08-20 1993-07-23 Hoya Corp メニスカス制御用平板
JP2007504816A (ja) * 2003-09-10 2007-03-08 オバチュアー リサーチ インコーポレイティド 細胞、胚または、卵母細胞を取り扱う装置と方法
JP2008532501A (ja) * 2005-03-10 2008-08-21 アレキサンダー，ロバート ウイルスの診断法およびこれに使用するウェル
JP2006280298A (ja) * 2005-04-01 2006-10-19 Nipro Corp 細胞培養用容器
JP2008045979A (ja) * 2006-08-15 2008-02-28 Yokogawa Electric Corp マイクロプレート
JP2010527582A (ja) * 2007-05-30 2010-08-19 株式会社ニコン 培養容器
JP2010200748A (ja) * 2009-02-09 2010-09-16 Dainippon Printing Co Ltd 細胞培養容器
JP2015047136A (ja) * 2013-09-03 2015-03-16 大日本印刷株式会社 細胞培養容器

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2017163378A1 (fr) * 2016-03-24 2017-09-28 株式会社ニコン Récipient de culture
CN112534038A (zh) * 2018-05-30 2021-03-19 太阳生物科学股份公司 一种用于培养生物材料的孔
CN112534038B (zh) * 2018-05-30 2024-07-05 太阳生物科学股份公司 一种用于培养生物材料的孔

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