WO2013179913A1 - Plateau de culture et kit associé - Google Patents

Plateau de culture et kit associé Download PDF

Info

Publication number
WO2013179913A1
WO2013179913A1 PCT/JP2013/063780 JP2013063780W WO2013179913A1 WO 2013179913 A1 WO2013179913 A1 WO 2013179913A1 JP 2013063780 W JP2013063780 W JP 2013063780W WO 2013179913 A1 WO2013179913 A1 WO 2013179913A1
Authority
WO
WIPO (PCT)
Prior art keywords
culture dish
culture
outer diameter
inner bottom
support member
Prior art date
Application number
PCT/JP2013/063780
Other languages
English (en)
Japanese (ja)
Inventor
陽子 淡海
益和 井家
誠 杉浦
裕子 諏訪
隆広 小笠原
Original Assignee
株式会社ジャパン・ティッシュ・エンジニアリング
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 株式会社ジャパン・ティッシュ・エンジニアリング filed Critical 株式会社ジャパン・ティッシュ・エンジニアリング
Priority to CN201380028128.2A priority Critical patent/CN104334713A/zh
Publication of WO2013179913A1 publication Critical patent/WO2013179913A1/fr

Links

Images

Classifications

    • 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
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
    • 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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • 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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/20—Material Coatings
    • 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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature

Definitions

  • the present invention relates to a culture dish and a culture kit.
  • a culture vessel used for culturing cells or tissues for example, a bottomed cylindrical culture dish having a diameter of 35 mm, 60 mm, 100 mm, 150 mm or the like is commercially available.
  • the cultured cells are cultured until they become full and the culture surface (inner bottom surface of the culture dish) is filled, and used as a final product for transplantation.
  • the larger the area of the culture surface the longer it takes to incubate to a confluent state and the more efficient the seeded cells are, so the area of the culture surface is smaller than the size of the required cell sheet. It is desirable not to be too big. For example, in the cornea, it is sufficient that a cell sheet having a diameter of about 25 mm is obtained. Therefore, the area of the culture surface is desirably about 25 mm.
  • the culture dish of Patent Document 1 has a problem in operability although it can cope with equipment designed to be compatible with general-purpose products. Specifically, there are problems that the opening of the cylinder is too small to take out the cell sheet, or that it is difficult to supply the culture solution into the cylinder.
  • the present invention has been made to solve such problems, and an object of the present invention is to provide a culture dish that is compatible with equipment designed to be compatible with general-purpose products and that has good operability. .
  • the culture dish and culture kit of the present invention employ the following means in order to achieve the above-mentioned main purpose.
  • the culture dish of the present invention is A dish body in which a bottomed cylindrical shape with an inner bottom surface serving as a culture surface and a small diameter narrowed portion and a cylindrical shape with a large diameter opening are connected via a connecting portion, A support member provided outside the bottom surface of the throttle portion; With The outer diameter of the opening and the outer diameter of the support member are the same as the outer diameter of a general-purpose culture dish.
  • the outer diameter of the opening and the outer diameter of the support member are the same as the outer diameter of the general-purpose culture dish, so that it can be used for equipment designed to be compatible with general-purpose products.
  • the culture is performed on the inner bottom surface of the small-diameter narrowed portion, a smaller cell sheet can be obtained than when the culture is performed using a general-purpose product.
  • the large-diameter opening is connected to the throttle part, the cell sheet is relatively easy to take out and the culture solution is easily supplied.
  • the support member is an annular support member formed so that an outer diameter thereof coincides with an outer diameter of the general-purpose culture dish, and a gap is formed between the support member and the throttle part. It is good as it is. In this way, the obtained cell sheet is transported in a state where it is immersed in the transport liquid for each culture dish, and the transport liquid is unlikely to remain on the support member when the culture dish is removed from the transport liquid and the cell sheet is operated. Therefore, the operation can be performed without worrying about dripping of the transport liquid.
  • the height of the culture dish may be the same as that of a general-purpose culture dish. In this way, it is easier to deal with general equipment and the like designed to fit a general-purpose culture dish.
  • the culture kit of the present invention comprises the culture dish of the present invention described above and a carrier placed on the inner bottom surface of the culture dish so that a part of the inner bottom surface is exposed.
  • the bottom surface is coated with a temperature-responsive polymer that switches between cell adhesion and cell detachability at a predetermined critical temperature, and the carrier has cell adhesion that does not depend on temperature.
  • the adhesion dependency when culturing adhesion-dependent cells, maintaining the inner bottom surface at a temperature at which cell adhesion is achieved, the adhesion dependency not only on the cell-adhesive carrier but also on the exposed inner bottom surface. Cells adhere and grow to form a cell sheet.
  • the cell sheet is detached from the inner bottom surface, and a cell sheet adhered only to the carrier is obtained.
  • the cell sheet is very thin and difficult to take out, in this culture kit, the cell sheet can be taken out simultaneously by holding the carrier and taking it out of the culture dish. Thus, culture and removal of adhesion-dependent cells can be performed easily.
  • the carrier is provided on the outer side of the annular part having an outer diameter slightly smaller than the inner diameter of the squeezed part of the culture dish, and the annular part is mounted on the culture dish. And an ear that rises along the side wall of the squeezed portion of the culture dish and partially protrudes above the side wall.
  • the ear part of the carrier protrudes above the side wall of the throttle part, it is easy to grasp the carrier and to take out the cell sheet more easily.
  • FIG. 1 is a perspective view of a culture dish 10.
  • FIG. It is the figure which looked at the culture dish 10 from the top. It is the figure which looked at the culture dish 10 from the bottom. It is a figure which shows the end surface in each cut surface of FIG. 2 is a perspective view of a culture kit 15.
  • FIG. It is explanatory drawing of the carrier 90.
  • FIG. It is explanatory drawing of the preparation method of the cell sheet using the culture kit.
  • FIG. It is explanatory drawing which shows the method of taking out a cell sheet from the culture dish.
  • FIG. 1 is a perspective view of the culture dish 10
  • FIG. 2 is a view of the culture dish 10 as viewed from above
  • FIG. 3 is a view of the culture dish 10 as viewed from below
  • FIG. It is. 4A corresponds to the AA section
  • FIG. 4B corresponds to the BB section
  • FIG. 4C corresponds to the CC section.
  • the culture dish 10 includes a dish body 20, a support member 60, and a bridging portion 70.
  • the culture dish 10 is made of polystyrene resin and is entirely transparent. Therefore, in FIGS. 1 to 3, the transparent back line should be described, but the transparent back line is omitted for convenience.
  • the dish body 20 includes a throttle part 30, an opening part 40, and a connecting part 50.
  • the dish body 20 has the same height as a general-purpose culture dish (here, 12 mm).
  • the narrowed portion 30 is formed in a bottomed cylindrical shape having an inner bottom surface 32 as a culture surface, and a leg 36 is provided on the bottom.
  • the diameter of the inner bottom surface 32 is ⁇ 25 mm here.
  • a temperature-responsive polymer is laid on the inner bottom surface 32.
  • the temperature-responsive polymer exhibits cell detachability at a temperature lower than a predetermined critical temperature (here, 32 ° C.) and cell adhesion at a higher temperature.
  • the temperature of the inner bottom surface 32 is maintained at, for example, about 37 ° C., and the inner bottom surface 32 is made cell-adhesive so that the cells adhere to the inner bottom surface 32 and proliferate. I will do it.
  • the temperature of the inner bottom surface 32 is lowered to, for example, about 20 ° C. to make the cells peelable, whereby the cells are detached from the inner bottom surface 32.
  • the temperature-responsive polymer is described in detail in, for example, Japanese Patent Application Laid-Open No. 2003-38170, and detailed description thereof is omitted here.
  • the legs 36 are intended to prevent scratches caused by the bottom of the plate body 20 coming into direct contact with the work table or the like, like the legs provided on the bottom of a general-purpose culture dish.
  • the legs 36 have the same outer diameter as the inner bottom surface 32 and are formed in an annular shape having a gap every 90 °.
  • the opening 40 is formed in a cylindrical shape having a diameter larger than that of the throttling portion 30 and has the same outer diameter as that of a general-purpose culture dish (here, ⁇ 35 mm).
  • the connecting part 50 connects the upper end of the throttle part 30 and the lower end of the opening part 40.
  • the connecting portion 50 has an arcuate shape in a sectional view that extends outward from the upper end of the side wall of the throttle portion 30 substantially horizontally and gradually extends upward.
  • the general-purpose culture dish means a commercially available product such as ⁇ 35 mm, ⁇ 60 mm, ⁇ 100 mm, and ⁇ 150 mm.
  • the same outer diameter and height as a general-purpose culture dish are the same as those that can be used for general equipment and the like designed to fit the general-purpose culture dish. That's fine.
  • the support member 60 is formed in an annular shape.
  • the support member 60 has the same outer diameter as that of a general-purpose culture dish (here, ⁇ 35 mm). Further, the lower end of the support member 60 is aligned with the lower end of the leg 36 of the dish body 20 in the height direction.
  • the support member 60 is provided with a protrusion 62.
  • the bridging part 70 is provided at four positions every 90 ° between the dish body 20 and the support member 60, and bridges the dish body 20 and the support member 60. Thereby, a gap 80 surrounded by the plate body 20, the support member 60, and the bridging portion 70 is formed.
  • the bridging portion 70 includes a first bridging portion 72 for bridging the narrowed portion 30 of the dish body 20 and the support member 60, and a second bridging portion 74 for bridging not only the throttling portion 30 but also the connecting portion 50. Are arranged alternately.
  • the bottom of the bridging portion 70 has the same position in the height direction as the bottom of the plate body 20 (excluding the legs 36).
  • FIG. 5 is a perspective view of the culture kit 15
  • FIG. 6 is an explanatory view of the carrier 90.
  • the culture kit 15 includes a culture dish 10 and a carrier 90 as shown in FIGS. Since the culture dish 10 has been described in “1. Culture dish” above, description thereof is omitted.
  • the carrier 90 is a sheet made of polyethylene terephthalate resin (PET) having a thickness of 0.1 mm or less (more preferably 0.05 mm or less), and the surface to be a cell adhesion surface is subjected to a hydrophilic treatment. Due to the hydrophilic treatment, it has cell adhesiveness independent of temperature.
  • “having cell adhesiveness independent of temperature” means that the cell adhesiveness is at least near the critical temperature (within plus or minus 10 ° C.) of the temperature-responsive polymer laid on the inner bottom surface 32 of the culture dish 10.
  • the carrier 90 includes an annular portion 92 having an outer diameter slightly smaller than the inner diameter of the squeezing portion 30 of the culture dish 10, and four ear portions 94 provided on the outer side of the annular portion 92 every 90 °. Yes.
  • the ear part 94 is used by being bent at a broken line part as shown in FIG.
  • a part of the ear part 94 protrudes above the rising side wall along the side wall of the throttle part 30 of the culture dish 10.
  • FIG. 7 is an explanatory diagram of a method for producing a cell sheet using the culture kit 15.
  • cells are dispersed from the tissue by trypsin treatment to prepare a cell suspension.
  • the carrier 90 is laid on the inner bottom surface 32 of the culture dish 10 (FIG. 7A). Then, feeder cells are laid on the inner bottom surface 32 on which the carrier 90 is laid, and a cell suspension is seeded (FIG. 7B).
  • As the feeder cells mouse fibroblasts whose proliferation ability is suppressed by inactivation treatment with mitomycin C or X-ray irradiation are used. After cell seeding, a DMEM / F12 mixed medium containing 10 v / v% FBS (fetal bovine serum) is supplied as a culture medium, and corneal cells are cultured in the presence of feeder cells.
  • the culture dish 10 is covered with a lid and cultured.
  • the culture dish 10 of the present embodiment has the same outer diameter of the opening 40 and the support member 60 as ⁇ 35 mm, so that the lid is designed to be compatible with a general-purpose culture dish of ⁇ 35 mm (general-purpose product). Can be applied as is.
  • cultivation may perform medium replacement
  • an automatic culture apparatus that automatically performs temperature control, medium replacement, or the like may be used.
  • Such devices are generally designed to be compatible with general-purpose culture dishes, and culture dishes that deviate from them are often unusable, and measures such as preparing a dedicated jig are required.
  • the culture dish 10 of the present embodiment can be directly applied to such an apparatus because the outer diameter of the opening 40 and the support member 60 is 35 mm, which is the same as that of a general-purpose culture dish.
  • the packaging container 100 is for wrapping the cell sheet together with the culture dish 10 for storage and transportation, and includes a container body 110, an adapter 170, and a lid 140.
  • the container body 110 is a container capable of storing a liquid such as a transport liquid, and is a cup-shaped container having a plurality of steps on the side wall. Specifically, a first step 114, a second step 116, and a third step 118 are formed upward from the inner bottom surface 112. A pair of vertical grooves 124 and 125 extending in the vertical direction are formed on the side wall of the container body 110. The pair of vertical grooves 124 and 125 are formed so as to face each other, and reach the second stepped portion 116 from the inner bottom surface 112 through the first stepped portion 114. On the outer peripheral side of the third step portion 118, a horizontal surface 120 is formed with a small step.
  • the adapter 170 is for holding the culture dish 10 in a state of being separated from the inner bottom surface 112 of the container main body 110.
  • the adapter 170 includes a cylinder 172 and a flange 174 provided at the upper end of the cylinder 172.
  • the cylinder 172 is a part for fixing the culture dish 10 inside thereof, and is formed so that the inner diameter is slightly larger than the outer diameter of the culture dish 10.
  • the cylinder 172 is provided with a claw 186 that is folded inward at the lower end so that the culture dish 10 does not shift downward.
  • the flange 174 is concavely curved and has a curved portion 176 whose plan view is a shape obtained by cutting a circle with two parallel strings, and a pair of vertical walls 178 rising vertically from the two strings. 179.
  • the outer periphery of the curved portion 176 has substantially the same shape as the inner periphery of the second step portion 116 of the container body 110.
  • a placement claw 188 having a horizontal bottom surface is provided so as to be stably placed on the upper surface of the first step portion 114 of the container body 110.
  • the vertical walls 178 and 179 are provided with distribution cutouts 180 and 181, respectively.
  • the adapter 170 includes a circulation hole 182 provided so as to penetrate both inside and outside the cylinder 172 and the flange 174, and a circulation slit 184 provided at the lower end of the cylinder 172.
  • the distribution slit 184 is adapted to catch the protrusion 62 provided on the support member 60 of the culture dish 10 and restricts the upward movement of the culture dish 10.
  • the lid 140 closes the opening of the container main body 110.
  • the lid body 140 includes a bottom surface 142 and a horizontal surface 144 provided on the outer peripheral side of the bottom surface 142, and the bottom surface 142 and the horizontal surface 144 are connected by a wall surface 146.
  • the lid body 140 includes a knob 148 raised from the bottom surface 142.
  • the bottom surface 142 is formed with a plurality of parallel reinforcing concave portions 150 that are convex downward.
  • the outer periphery of the lid 140 has substantially the same shape as the outer periphery of the third step portion 118 of the container body 110.
  • the cell sheet is packaged together with the culture dish 10 as follows. First, the produced cell sheet is washed with a washing solution a plurality of times. Subsequently, the culture dish 10 with the cell sheet is pushed into the cylinder 172 of the adapter 170 and fixed, and the adapter 170 is set in the container body 110.
  • the adapter 170 is set on the container main body 110, the cylinder 172 enters between the inner bottom surface 112 of the container main body 110 and the first stepped portion 114, the curved portion 176 of the flange 174 enters the inner periphery of the second stepped portion 116, A placement claw 188 provided on the bottom surface of the bending portion 176 is placed on the first step portion 114.
  • the pair of vertical walls 178 and 179 are opposed to the pair of vertical grooves 124 and 125 provided in the container body 110, and the upper region and the lower side of the adapter 170 are interposed between the adapter 170 and the container body 110.
  • the transport liquid is injected into the region below the adapter 170 from the passage formed between the vertical walls 178 and 179 and the vertical grooves 124 and 125, and the water level of the second stepped portion 116 of the container body 110. Inject until the top level is reached.
  • the transport liquid also enters the region above the adapter 170 through the distribution notches 180 and 181.
  • an air pool may be formed below the culture dish 10, but such air is not limited to the passage formed between the vertical walls 178 and 179 and the vertical grooves 124 and 125, and the adapter 170.
  • the adapter 170 through the distribution hole 182 and the distribution slit 184, the gap provided between the legs 36 and 36 of the culture dish 10, and the gap 80 between the dish body 20 and the support member 60. It can be pulled upward.
  • the lid 140 is capped.
  • a sealing sheet 210 (indicated by hatching in the drawing) is placed above the packaging container 100 and thermally fused to the upper surface of the packaging container 100, thereby obtaining a package 200 as shown in FIG.
  • the sealing sheet 210 includes an ear 212 that becomes a handle when the sealing sheet 210 is peeled off from the packaging container 100.
  • the sealing sheet 210 of the packaging body 200 is peeled off from the packaging container 100, the lid 140 is removed from the container body 110, and the adapter 170 is removed from the container body 110. Then, by tilting the culture dish 10 while being fixed to the adapter 170, unnecessary transport liquid in the culture dish 10 is discharged from the distribution cutouts 180 and 181. In addition, about discharge
  • the cell sheet is taken out from the culture dish 10. Subsequent operations will be described with reference to FIG. First, by cooling the culture dish 10 to about 20 ° C., the inner bottom surface 32 of the culture dish 10 is made cell-peelable, and the part where the inner bottom surface 32 and the cell sheet are in direct contact is peeled off. At this time, the cell sheet remains adhered to the carrier 90 which is a cell adhesive sheet having no temperature responsiveness. Next, the ear
  • a scalpel FIG.10 (B)
  • the outer diameter of the opening 40 and the outer diameter of the support member 60 are the same as the outer diameter of a general-purpose culture dish, so that it is designed to be compatible with a general-purpose product having a diameter of 35 mm. It can correspond to the equipment.
  • equipment designed to be compatible with a general-purpose culture dish having a diameter of 35 mm can be used as it is.
  • the culture is performed on the inner bottom surface 32 having a diameter of 25 mm, a smaller cell sheet can be obtained than when the culture is performed using a general-purpose product having a diameter of 35 mm, and the following advantages are obtained.
  • a cell sheet of about ⁇ 25 mm is sufficient, so when culturing with a general-purpose product of ⁇ 35 mm, it takes time to reach a confluent state, many cells are seeded, and extra cells A cell sheet may be formed in the region.
  • the culture dish 10 is used, a cell sheet can be obtained efficiently.
  • the large-diameter opening 40 is connected to the throttle portion 30 having a bottomed cylindrical shape, it is relatively easy to take out the cell sheet. In this regard, it seems that a cell sheet can be easily taken out even in a bottomed cylindrical culture vessel having no opening 40 if the depth of the vessel is shallow. However, it is difficult to make the container shallower because cell culture requires a certain amount of the culture medium, and the container must have sufficient depth to prevent the culture medium from spilling during operation. . Further, if the culture solution is supplied to the large-diameter opening 40, the culture solution inevitably flows into the small-diameter throttle portion 30, so that the culture solution can be easily supplied to the culture surface.
  • the support member 60 is an annular support member formed so as to have the same outer diameter as that of a general-purpose culture dish, and a gap 80 is formed between the throttling part 30, the obtained cells
  • the transport liquid hardly remains on the support member 60. Therefore, the operation can be performed without worrying about dripping of the transport liquid.
  • the dish body 20 has the same height as a general-purpose culture dish, and the culture dish 10 has the same height as a general-purpose culture dish as a whole. For this reason, it is easier to deal with general equipment and the like designed to fit a general-purpose culture dish.
  • the inner bottom surface 32 of the culture dish 10 is laid with a temperature-responsive polymer that switches between cell adhesion and cell detachability at a predetermined critical temperature, and further has a cell adhesion that does not depend on temperature. Then, a carrier 90 having such a shape that a part of the inner bottom surface 32 is exposed is placed on the inner bottom surface 32 and used. For this reason, it is easy to culture and take out the cell sheet.
  • the carrier 90 since the carrier 90 includes an annular portion 92 having an outer diameter slightly smaller than the inner diameter of the narrowed portion 30 of the culture dish 10, the cell sheet can be taken out with the entire outer periphery adhered to the carrier 90. . For this reason, shrinkage or deformation of the cell sheet can be suppressed. Moreover, since the ear
  • the culture dish 10 can be packaged and stored or transported.
  • a general-purpose culture dish can be used instead of the culture dish 10.
  • the support member 60 is an annular support member, and the gap 80 is formed between the support member 60 and the throttle unit 30, but the outer diameter is the same as the outer diameter of a general-purpose culture dish. If it is, it will not be limited to this.
  • the support member 60 may be a disc shape in which the gap 80 is not formed between the support portion 30 and a support member 60 may be formed of a plurality of rod-shaped members that extend radially from the stop portion 30. Even in this case, it is possible to cope with equipment designed to be compatible with general-purpose products.
  • the culture dish 10 is made of polystyrene resin, but is not limited thereto.
  • various resins such as polyvinyl chloride resin, polyethylene resin, acrylic resin, polypropylene resin, polymethylpentene resin, fluorine resin, phenol resin, urea resin, epoxy resin, melamine resin, and silicon resin can be used.
  • the culture dish 10 shall be the whole transparent so that it may be easy to confirm the state of a cell, it is not limited to this. For example, if the bottom of the diaphragm portion 30 is transparent, microscopic observation is possible, and the other portions may not be transparent.
  • the outer diameter of the opening 40 and the support member 60 is 35 mm, the diameter of the inner bottom surface 32 is 25 mm, and the height is 12 mm.
  • the present invention is not limited to this.
  • the diameter of the inner bottom surface 32 may be ⁇ 10 mm.
  • the diameter of the inner bottom surface 32 may be set to 40 mm.
  • the opening 40 and the support member 60 can be used for equipment compatible with a general-purpose product having a diameter of 60 mm.
  • the outer diameter can be set to 60 mm.
  • the height may be the same as that of the general-purpose product or may be different.
  • the inner bottom surface 32 is provided with a temperature-responsive polymer, but is not limited thereto.
  • a cell-adhesive property may be simply provided by laying a cell-adhesive polymer or surface treatment such as hydrophilic treatment.
  • adhesion-dependent cells such as corneal cells can be cultured.
  • the cell sheet can be peeled off by performing an enzyme treatment to peel off the cells.
  • the surface treatment may be any treatment as long as cell adhesion can be obtained.
  • coating with a chemical material such as an adhesive factor such as collagen or fibronectin or a temperature-responsive polymer, ion irradiation, plasma treatment, etc. Surface modification etc. are mentioned.
  • bridging portions 70 are provided every 90 °, but the present invention is not limited to this. For example, it may not be provided at equal intervals, or may be provided at 3 or less locations or at 5 or more locations.
  • crosslinking part 72 which bridge
  • crosslinking part 74 which bridge
  • the first bridging part 72 has an advantage that liquid does not easily stay at the bottom of the culture dish 10, and the second bridging part 74 has an advantage that the strength is easily increased. do it.
  • bridging part 70 shall have the same position of the height direction as the bottom (except the leg 36) of the plate main body 20, it is not limited to this. For example, the lower end of the leg 36 and the position in the height direction may be the same.
  • the legs 36 are provided on the bottom of the dish body 20, but the legs 36 may not be provided.
  • the leg 36 has a gap every 90 °.
  • the leg 36 may not have a gap, the gap may not be provided at equal intervals, and the gap interval may not be 90 °. Also good.
  • the shape of the connecting portion 50 is a circular arc shape in a cross-sectional view that extends outward from the upper end of the side wall of the throttle portion 30 substantially horizontally and gradually extends upward, but is not limited thereto.
  • a gentle slope shape may be formed from the upper end of the side wall of the throttle part 30 toward the upper end of the opening part 40, or a gentle slope shape may be formed between the upper end of the side wall of the throttle part 30 and the upper end of the opening part 40. It is good also as the shape made to stand perpendicularly
  • corneal cells were cultured, but the cells to be cultured are not limited to this.
  • it can be suitably used for culturing adhesion-dependent cells such as skin and oral mucosa.
  • the carrier 90 is laid and used, but the carrier 90 is not necessarily laid.
  • the carrier 90 has the annular portion 92 having an outer diameter slightly smaller than the inner diameter of the narrowed portion 30 of the culture dish 10.
  • the carrier 90 is not limited to this, and the carrier 90 is attached to the culture dish 10. What is necessary is just to expose a part of the inner surface 32 when it is placed on the inner bottom surface 32.
  • the carrier 90 is provided with the four ears 94 for each 90 ° of the ears 94, but the ears 94 may not be provided or may not be provided at regular intervals. Three or less or five or more may be provided.
  • the obtained cell sheet is packaged in the packaging container 100 and then taken out and used, but such packaging may not be performed. Moreover, you may package with packaging containers other than the packaging container 100. FIG.
  • the present invention can be used in a cell culture technique for culturing cells in vitro.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Sustainable Development (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Clinical Laboratory Science (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Physics & Mathematics (AREA)
  • Thermal Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

Cette invention concerne un plateau de culture (10) comprenant un corps de plateau (20) et un élément de support (60). Le corps de plateau (20) comprend : une partie resserrée (30) de forme cylindrique, à extrémité fermée ayant une surface de fond intérieure (32) servant de surface de culture; une partie ouverture (40) de forme cylindrique et de grand diamètre; et une partie raccordement (50) pour raccorder la partie resserrée (30) et la partie ouverture (40). La partie ouverture (40) a le même diamètre extérieur qu'un plateau de culture à caractère général. L'élément de support (60) revêtit une forme annulaire et a le même diamètre extérieur que le plateau de culture à caractère général, et le plateau de culture (10)a la même hauteur que le plateau de culture à caractère général.
PCT/JP2013/063780 2012-05-31 2013-05-17 Plateau de culture et kit associé WO2013179913A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201380028128.2A CN104334713A (zh) 2012-05-31 2013-05-17 培养皿及培养套件

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2012124818A JP5990403B2 (ja) 2012-05-31 2012-05-31 培養皿及び培養キット
JP2012-124818 2012-05-31

Publications (1)

Publication Number Publication Date
WO2013179913A1 true WO2013179913A1 (fr) 2013-12-05

Family

ID=49673116

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2013/063780 WO2013179913A1 (fr) 2012-05-31 2013-05-17 Plateau de culture et kit associé

Country Status (3)

Country Link
JP (1) JP5990403B2 (fr)
CN (1) CN104334713A (fr)
WO (1) WO2013179913A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015076115A1 (fr) * 2013-11-22 2015-05-28 株式会社ジャパン・ティッシュ・エンジニアリング Récipient de culture de cellules et récipient dans lequel une culture de cellule a été logée
CN110511871A (zh) * 2019-09-25 2019-11-29 西北农林科技大学 一种细胞共培养的装置及牛肌细胞与脂肪细胞的共培养方法
US20200100775A1 (en) * 2017-06-05 2020-04-02 Orthocell Limited A medical procedure kit and a related method
CN117887561A (zh) * 2023-12-22 2024-04-16 中科院南昌高新技术产业协同创新研究院 一种无细胞蛋白质培养皿及培养方法

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP6307686B2 (ja) * 2012-12-05 2018-04-11 三星ディスプレイ株式會社Ｓａｍｓｕｎｇ Ｄｉｓｐｌａｙ Ｃｏ．，Ｌｔｄ． アミン誘導体、有機発光材料及びそれを用いた有機エレクトロルミネッセンス素子
JP2016013111A (ja) * 2014-07-03 2016-01-28 大日本印刷株式会社 底面部を剥離可能な培養容器
JP7278523B2 (ja) * 2017-12-26 2023-05-22 株式会社幹細胞＆デバイス研究所 細胞培養容器
CN108913600B (zh) * 2018-08-16 2023-06-27 杭州德适生物科技有限公司 一种显微授精操作皿

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000342245A (ja) * 1999-06-01 2000-12-12 Yuzo Yamaguchi 培養菌選別装置
JP2001224355A (ja) * 2000-05-25 2001-08-21 Kyodo Seihin Kk 両面シャーレ
WO2002008387A1 (fr) * 2000-07-21 2002-01-31 Cellseed Inc. Feuille cellulaire du type muscle cardiaque, construction tridimensionnelle, tissu du type muscle cardiaque et procede de production associe
JP2005333823A (ja) * 2004-05-24 2005-12-08 Olympus Corp 培養容器用アダプタおよび培養処理装置
JP2006280298A (ja) * 2005-04-01 2006-10-19 Nipro Corp 細胞培養用容器
JP2007229250A (ja) * 2006-03-01 2007-09-13 Olympus Corp 細胞シートのリリース方法
JP3145813U (ja) * 2008-08-08 2008-10-23 株式会社ベルテックス シャーレ
WO2008143149A1 (fr) * 2007-05-11 2008-11-27 Dai Nippon Printing Co., Ltd. Feuille de cellules ayant une bonne stabilité dimensionnelle, son procédé de fabrication, et support de culture cellulaire pour une utilisation dans le procédé

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1753696B (zh) * 2003-02-20 2010-04-28 创革医疗科技有限公司 再生角膜内皮细胞片、制造方法及其使用方法

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000342245A (ja) * 1999-06-01 2000-12-12 Yuzo Yamaguchi 培養菌選別装置
JP2001224355A (ja) * 2000-05-25 2001-08-21 Kyodo Seihin Kk 両面シャーレ
WO2002008387A1 (fr) * 2000-07-21 2002-01-31 Cellseed Inc. Feuille cellulaire du type muscle cardiaque, construction tridimensionnelle, tissu du type muscle cardiaque et procede de production associe
JP2005333823A (ja) * 2004-05-24 2005-12-08 Olympus Corp 培養容器用アダプタおよび培養処理装置
JP2006280298A (ja) * 2005-04-01 2006-10-19 Nipro Corp 細胞培養用容器
JP2007229250A (ja) * 2006-03-01 2007-09-13 Olympus Corp 細胞シートのリリース方法
WO2008143149A1 (fr) * 2007-05-11 2008-11-27 Dai Nippon Printing Co., Ltd. Feuille de cellules ayant une bonne stabilité dimensionnelle, son procédé de fabrication, et support de culture cellulaire pour une utilisation dans le procédé
JP3145813U (ja) * 2008-08-08 2008-10-23 株式会社ベルテックス シャーレ

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2015076115A1 (fr) * 2013-11-22 2015-05-28 株式会社ジャパン・ティッシュ・エンジニアリング Récipient de culture de cellules et récipient dans lequel une culture de cellule a été logée
US10370631B2 (en) 2013-11-22 2019-08-06 Japan Tissue Engineering Co., Ltd. Cell culture package and package containing cell-culture material
US20200100775A1 (en) * 2017-06-05 2020-04-02 Orthocell Limited A medical procedure kit and a related method
CN110511871A (zh) * 2019-09-25 2019-11-29 西北农林科技大学 一种细胞共培养的装置及牛肌细胞与脂肪细胞的共培养方法
CN117887561A (zh) * 2023-12-22 2024-04-16 中科院南昌高新技术产业协同创新研究院 一种无细胞蛋白质培养皿及培养方法

Also Published As

Publication number Publication date
CN104334713A (zh) 2015-02-04
JP5990403B2 (ja) 2016-09-14
JP2013247909A (ja) 2013-12-12

Similar Documents

Publication Publication Date Title
JP5990403B2 (ja) 培養皿及び培養キット
JP6711825B2 (ja) 培養容器輸送用セットおよび細胞・生体組織輸送ユニット
JP5674953B2 (ja) 細胞培養インサート
US7022518B1 (en) Apparatus and method for co-culturing of cells
JP2013138684A (ja) バイオリアクタの動作方法
JP6431341B2 (ja) 接着性細胞の培養方法
JP6642017B2 (ja) バイオロジカルインジケータ用基材、バイオロジカルインジケータ、バイオロジカルインジケータ用基材保管体、バイオロジカルインジケータ保管体、バイオロジカルインジケータ用基材保管体の製造方法、バイオロジカルインジケータ保管体の製造方法および殺菌装置の殺菌能力評価方法
US20190185799A1 (en) Container for transporting cells and biotissue
JP6459219B2 (ja) 細胞培養容器
JP2009089715A (ja) 培養細胞シート包装体及びシート挟持部材
WO2016117281A1 (fr) Récipient de culture cellulaire, procédé de culture cellulaire et procédé d'utilisation du récipient de culture cellulaire
JP2013099278A (ja) 細胞培養容器の製造方法
WO2005037986A1 (fr) Emballage pour feuille de cellules en culture
US20120058560A1 (en) Bioreactor System
JP2023101552A (ja) 挟持具
JP7039202B2 (ja) 液体受容空間を具備する脆弱物保持デバイス
JP2016136871A (ja) 異なる孔径の２種類の多孔膜を有する細胞培養基材、及び該細胞培養基材を有する細胞培養器具
JP2014168403A (ja) 細胞培養容器
JP6983562B2 (ja) 液体保持空間を具備する脆弱物保持デバイス
JP2019017343A (ja) ガイド機構を具備する脆弱物保持デバイス
JP7181200B2 (ja) 脆弱物保持デバイス
JP2005318868A (ja) 培養容器
WO2017078007A1 (fr) Récipient de culture
JP2014168402A (ja) 細胞培養容器
JP7353086B2 (ja) 閉鎖系細胞培養容器

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 13797783

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 13797783

Country of ref document: EP

Kind code of ref document: A1