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WO2021132480A1 - Substrat de culture et boîte de culture - Google Patents

Substrat de culture et boîte de culture Download PDF

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Publication number
WO2021132480A1
WO2021132480A1 PCT/JP2020/048493 JP2020048493W WO2021132480A1 WO 2021132480 A1 WO2021132480 A1 WO 2021132480A1 JP 2020048493 W JP2020048493 W JP 2020048493W WO 2021132480 A1 WO2021132480 A1 WO 2021132480A1
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WO
WIPO (PCT)
Prior art keywords
culture
culture medium
less
microwells
shape
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Ceased
Application number
PCT/JP2020/048493
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English (en)
Japanese (ja)
Inventor
千佳 土屋
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
AGC Inc
Original Assignee
Asahi Glass Co Ltd
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 Asahi Glass Co Ltd filed Critical Asahi Glass Co Ltd
Priority to JP2021567624A priority Critical patent/JPWO2021132480A1/ja
Publication of WO2021132480A1 publication Critical patent/WO2021132480A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • C12M3/00—Tissue, human, animal or plant cell, or virus culture apparatus

Definitions

  • the present invention relates to a culture medium and a culture container.
  • Spheroid culture is well known in which cells derived from humans and animals are artificially cultured in a culture vessel or the like and aggregated three-dimensionally. It is known that in spheroid culture, cell populations form a three-dimensional structure and cells interact with each other, so that they are closer to the three-dimensional structure in vivo and exhibit superior characteristics compared to planar adhesive culture. ing. In fact, spheroid culture is often used for anti-cancer drug screening using cancer cells and for proliferation and differentiation of pluripotent stem cells.
  • the opening area expands as it approaches the opening end, a recess for accommodating cells and a culture solution is provided at the bottom of the container body, and a plurality of microwells for assembling cells by gravity are provided in the recess.
  • the culture vessel provided on the bottom surface of the above is disclosed. If the culture vessel is not flat, it will be difficult to focus the cells, and it will be easily affected by external light, making it difficult to observe the cells. Therefore, it is desired that the culture vessel has excellent flatness.
  • Patent Document 2 in a cell culture plate in which a sheet having microwells is attached to the back surface of a well plate having through holes to form the bottom surface of the wells, injection-molded pingates are arranged at specific positions. It is disclosed to improve the flatness of the.
  • Patent Document 2 in a culture vessel in which a base material having a plurality of microwells is attached to a frame body, even if the frame body is highly flat, the adhesiveness between the base material and the frame body is lowered, or cells are formed. May be difficult to observe.
  • An object of the present invention is to provide a culture medium having excellent adhesion between the culture medium and the frame and easy observation of cells and the like, and a culture container provided with the culture medium.
  • the present invention has the following aspects.
  • [4] The culture medium according to any one of [1] to [3], wherein the plurality of microwells are formed by unevenness formed of a curved surface.
  • [5] The culture medium according to any one of [1] to [4], wherein the average number of microwells per unit area is 10 pieces / cm 2 or more.
  • [6] The culture medium according to any one of [1] to [5], wherein the material of the culture medium is synthetic resin or glass.
  • [7] The culture medium according to any one of [1] to [6], wherein the average thickness of the culture medium is 50 ⁇ m or more and 2000 ⁇ m or less.
  • the present invention it is possible to provide a culture medium having excellent adhesion between the culture medium and the frame and observing cells easily, and a culture container provided with the culture medium.
  • FIG. 3 is a cross-sectional view taken along the line AA of the culture vessel of FIG. It is sectional drawing which showed an example of the plurality of microwells of a culture area.
  • the “culture region” means a region on the surface of the culture substrate that constitutes the bottom surface of the portion in which the cells of the culture vessel are cultured and has a plurality of microwells.
  • a “microwell” is a depression having an opening diameter of 2000 ⁇ m or less and a depth of 2000 ⁇ m or less.
  • a “continuous curved surface” is a surface that does not include a flat surface.
  • Microwell opening diameter is the diameter of the microwell opening in the reference plane. When the opening shape of the microwell on the reference plane in a plan view is not a perfect circle, the diameter of the circumscribed circle of the opening shape is defined as the diameter of the opening of the microwell.
  • the “microwell depth” is the distance between the deepest part of the microwell and the reference plane.
  • the “reference surface of the culture medium” is a surface including the outer peripheral edge of the surface including the culture region of the culture medium. For example, when a plurality of microwells are formed on the surface of the culture substrate by laser irradiation, the surface of the culture substrate before laser irradiation coincides with the reference plane. When the culture region is partially provided on a part of the surface of the culture substrate, the surface of the region other than the culture region among the surfaces including the culture region of the culture substrate coincides with the reference plane.
  • the "frame body” is a member that constitutes a culture container together with a culture base material, and is a member that includes a side wall portion that surrounds the culture area with the culture base material attached to the bottom.
  • the culture substrate of the present invention is a plate-shaped substrate, and is a culture region in which at least a part of one surface has a plurality of microwells.
  • the region constituting the bottom surface of the portion of the culture medium in which the cells are cultured is formed on the surface facing upward of the culture substrate. It becomes a culture area.
  • the culture region may be partially formed on the surface of the culture substrate or may be formed on the entire surface of the culture substrate.
  • the entire surface 10a facing upward of the culture base 10 is cultured. It becomes the area 12.
  • the surface of the culture base 10 facing upward is the culture region 12.
  • the culture substrate may be attached to the lower side of the frame without the bottom surface 112a.
  • the amount of warpage of both surfaces of the culture substrate is 400 ⁇ m or less. That is, both the amount of warpage of the surface of the culture substrate including the culture region and the amount of warpage of the surface opposite to the surface including the culture region are 400 ⁇ m or less.
  • the amount of warpage of the entire surface of the culture base material is not more than the above upper limit value, the adhesiveness between the culture base material and the frame is excellent.
  • the flatness of the culture vessel provided with the culture substrate becomes excellent, the cells can be easily focused, and the cells are less likely to be affected by external light, so that the cells can be easily observed.
  • the amount of warpage of both surfaces of the culture substrate is preferably 380 ⁇ m or less, more preferably 350 ⁇ m or less.
  • the amount of warpage of the entire surface of the culture substrate is the difference between the maximum value and the minimum value of the amount of warpage measured by a non-contact shape measurement system using a laser displacement meter.
  • the ratio of the total area of the culture area to the total area of the surface including the culture area of the culture substrate is preferably 10% or more and 100% or less, and more preferably 30% or more and 90% or less.
  • the ratio is equal to or higher than the lower limit of the above range, spheroids having a desired size are likely to be formed.
  • the ratio is not more than the upper limit of the above range, the culture medium is less likely to be distorted, and the adhesiveness to the frame is improved.
  • the total area of the surface including the culture region of the culture substrate is preferably 5 cm 2 or more and 700 cm 2 or less.
  • the culture substrate is used in the form of a microplate, it is 80 cm 2 or more and 120 cm 2 or less, when it is used in the dish shape, it is 9 cm 2 or more and 150 cm 2 or less, and when it is used in the flask shape, it is 25 cm 2 or more and 700 cm 2 or less, large.
  • 250 cm 2 or more and 700 cm 2 or less per stage is more preferable.
  • the total area of the surface including the culture region is equal to or more than the lower limit of the above range, the entire bottom surface of the frame can be covered, and the adhesiveness with the frame is improved.
  • the total area of the surface including the culture region is not more than the upper limit of the above range, the culture substrate is less likely to be distorted and the adhesiveness to the frame is improved.
  • the total area of the culture regions is the total area of the plurality of culture regions.
  • the plan-view shape of the culture substrate can be appropriately set according to the shape of the frame to be attached, and examples thereof include a round shape and a rectangular shape.
  • the average thickness of the culture medium is preferably 50 ⁇ m or more and 2000 ⁇ m or less.
  • the average thickness is more preferably 100 ⁇ m or more and 250 ⁇ m or less, and further preferably 130 ⁇ m or more and 200 ⁇ m or less.
  • the culture substrate is a resin, it is more preferably 300 ⁇ m or more and 1800 ⁇ m or less, further preferably 500 ⁇ m or more and 1500 ⁇ m or less, and even more preferably 810 ⁇ m or more and 1000 ⁇ m or less.
  • the average thickness of the culture base material is within the above range, the deformation of the culture base material due to the formation of microwells can be reduced.
  • the culture area of the culture medium has a plurality of microwells. By having a plurality of microwells in the culture region, during cell culture, cells fall into the microwells and spheroids are formed in the wells.
  • the plurality of microwells 14 included in the culture region 12 of the culture substrate 10 are formed by irregularities formed of curved surfaces.
  • the plurality of microwells are composed of irregularities composed of continuous curved surfaces. If the inside of the microwell is a curved surface, a beautiful spherical spheroid is likely to be formed.
  • the boundary surface between the microwells is a curved surface, cells are less likely to remain on the boundary surface and easily fall into the microwells.
  • the boundary surface between the microwells may be flat.
  • the dotted line indicates the reference plane of the culture medium 10. This reference plane is a plane including the outer peripheral edge of the surface including the culture region 12 of the culture substrate 10.
  • a plurality of microwells composed of irregularities formed of curved surfaces can be formed by, for example, laser irradiation.
  • the opening shape of the microwell in a plan view can be appropriately selected according to the type of cells to be cultured and the size of the spheroid to be formed, and examples thereof include a round shape, an elliptical shape, a polygonal shape, and a donut shape. ..
  • the formation pattern of a plurality of microwells can be appropriately selected according to the type of cells to be cultured and the shape of the frame to be attached, and examples thereof include a lattice shape, a polygonal shape, a honeycomb shape, a round shape, and a donut shape.
  • the microwell formation pattern may be adjusted according to the opening shape of the side wall portion in a plan view. For example, in the case of a microplate-shaped culture vessel in which the opening shape of the side wall portion of the frame body is round in the plan view, the formation pattern can be made round.
  • the average diameter d (FIG. 5) of the opening of the microwell can be appropriately adjusted according to the desired size of the spheroid, and is preferably 20 ⁇ m or more and 1500 ⁇ m or less, more preferably 100 ⁇ m or more and 1000 ⁇ m or less, and further preferably 200 ⁇ m or more and 800 ⁇ m or less.
  • a spheroid having a desired size is likely to be formed.
  • the average diameter d of the opening of the microwell is equal to or less than the upper limit of the above range, it is easy to prevent cells from spilling from the microwell, and it is easy to form a spheroid having a uniform size.
  • the diameter of the opening of the microwell is measured by a laser microscope (manufactured by KEYENCE CORPORATION) or the like.
  • the average depth D (FIG. 5) of the microwell is preferably 10 ⁇ m or more and 1500 ⁇ m or less, more preferably 50 ⁇ m or more and 1000 ⁇ m or less, and further preferably 100 ⁇ m or more and 600 ⁇ m or less.
  • the average depth D of the microwells is equal to or greater than the lower limit of the above range, spheroids having a desired size are likely to be formed.
  • the average depth D of the microwells is equal to or less than the upper limit of the above range, it is easy to prevent cells from spilling from the microwells, and it is easy to form spheroids having a uniform size.
  • the depth of the microwell is measured by a laser microscope (manufactured by KEYENCE CORPORATION) or the like.
  • the opening area of the microwell is preferably 0.15 mm 2 or more 0.50 mm 2 or less, more preferably 0.20 mm 2 or more 0.35 mm 2 or less, more preferably 0.24 mm 2 or more 0.30 mm 2 or less.
  • the opening area of the microwell is equal to or larger than the lower limit of the above range, spheroids having a desired size are likely to be formed.
  • the opening area of the microwell is equal to or less than the upper limit of the above range, it is easy to prevent cells from spilling from the microwell, and it is easy to form a spheroid having a uniform size.
  • the opening area of the microwell is measured by a laser microscope (manufactured by KEYENCE CORPORATION) or the like.
  • the average number of microwells per unit area in the culture region is preferably 10 pieces / cm 2 or more, more preferably 15 pieces / cm 2 or more, and even more preferably 20 pieces / cm 2 or more.
  • the average number of microwells per unit area in the culture region is preferably 10,000 pcs / cm 2 or less, more preferably 5,000 pcs / cm 2 or less, and even more preferably 1000 pcs / cm 2 or less.
  • spheroids having a desired size are likely to be formed.
  • synthetic resin or glass is preferable.
  • the synthetic resin constituting the culture substrate include acrylic resin, polystyrene resin, polyester resin, polycarbonate resin, polypropylene resin, silicone resin, polyethylene terephthalate (PET) resin, vinyl chloride, high-density polyethylene, polyether sulfan, and the like.
  • PET copolymers include PET copolymers, polypropylene resins, Permanox TM, cycloolefin polymer resins, Cytop TM, and the like.
  • a silicone resin is used as the culture substrate, it is easy to prevent cells from adhering to the surface of the culture substrate.
  • the synthetic resin constituting the culture substrate may be one kind or two or more kinds.
  • Synthetic resins include substances that suppress cell adhesion (eg, phospholipid polymers (eg 2-methacryloyloxyethyl phosphorylcholine), polyhydroxyethyl methacrylate, fluorine-containing compounds, polyethylene glycol, etc.) and colorants (eg, oxidation). Titanium, carbon, etc.) may be blended.
  • Examples of the glass constituting the culture medium include quartz glass, borosilicate glass, phosphate glass, and chemically strengthened glass.
  • the glass constituting the culture substrate may be one kind or two or more kinds.
  • a cell adhesion inhibitor may be applied to the culture region on the surface of the culture substrate to form a film.
  • the coating film By forming the coating film on the surface of the culture region including the inner surface of the microwell, it is possible to prevent cells from adhering to the surface of the culture region. Therefore, it becomes easy for cells to aggregate in the microwell to form a spheroid, and it becomes easy to take out the spheroid from the microwell.
  • the cell adhesion inhibitor examples include phospholipid polymers (2-methacryloyloxyethyl phosphorylcholine, etc.), polyhydroxyethyl methacrylate, fluorine-containing compounds, and polyethylene glycol.
  • phospholipid polymers (2-methacryloyloxyethyl phosphorylcholine, etc.)
  • polyhydroxyethyl methacrylate examples include polyethylene glycol.
  • the method for producing the culture medium is not particularly limited. For example, a method of molding using a mold having a cavity surface having a shape complementary to the surface shape of a culture region having a plurality of microwells, and a process of forming a plurality of microwells on the surface of a molded base material.
  • the method of applying can be exemplified.
  • Examples of the method for forming a plurality of microwells after molding include laser irradiation (CO 2 laser, YAG laser, excimer laser, etc.), nanoimprint, press, and the like.
  • CO 2 laser laser irradiation
  • YAG laser YAG laser
  • excimer laser etc.
  • nanoimprint press, and the like.
  • the culture substrate is sandwiched (loaded) between a pair of flattening plates and heated.
  • an annealing treatment in which the culture substrate is sandwiched (loaded) between a pair of flattening plates and heated.
  • the culture medium itself is easily distorted, but by performing the annealing treatment, it becomes easy to obtain a culture medium having a warp amount of 400 ⁇ m or less on both surfaces.
  • the material of the flattening plate include stainless steel (SUS) and quartz glass.
  • the load condition (weight stone condition) for the annealing treatment is preferably 100 g or more and 2000 g or less, and more preferably 500 g or more and 1200 g or less.
  • the warp of the culture medium can be uniformly reduced.
  • the weight stone it is preferable to use a weight stone having high flatness (warp amount ⁇ 5 ⁇ m) so that the load is evenly applied.
  • the heating conditions for the annealing treatment are preferably 80 ° C. or higher and 100 ° C. or lower, and more preferably 80 ° C. or higher and 90 ° C. or lower.
  • the heating conditions are equal to or higher than the lower limit of the above range, the stress in the culture medium can be relaxed and the warpage can be reduced.
  • the heating conditions are not more than the upper limit of the above range, deformation due to melting of the culture substrate can be suppressed.
  • the culture vessel of the present invention includes the culture medium of the present invention and a frame.
  • the culture base material of the present invention is attached to the bottom of the frame, so that the culture region on the surface of the culture base material becomes the bottom surface of the portion for culturing cells.
  • Examples of the container shape of the culture vessel of the present invention include a flask shape, a petri dish shape, a microplate shape, and a multilayer flask shape.
  • the shape of the culture vessel is preferably one of a flask shape, a petri dish shape, a microplate shape, and a multilayer flask shape from the viewpoint that a beautiful spherical spheroid is easily formed.
  • the shape of the culture vessel can be adjusted by the shape of the frame.
  • Examples of the petri dish-shaped culture container include the culture container 100 illustrated in FIG. In the culture container 100, the culture base material 10 is attached to the bottom surface 112a of the petri dish-shaped frame 110 so that the culture region 12 faces upward.
  • the frame body 110 includes a bottom portion 112 and a side wall portion 114 rising from the peripheral edge portion of the bottom portion 112.
  • the entire surface 10a of the culture base material 10 is the culture region 12, and the culture region 12 is surrounded by the side wall portion 114 of the frame body 110.
  • microplate-shaped culture container examples include the culture container 200 illustrated in FIGS. 3 and 4.
  • the culture base material 10 is attached to the bottom of the frame body 210 having a plurality of through holes 212.
  • the frame body 210 includes a flat plate portion 214 having a rectangular shape in a plan view, and a plurality of tubular portions 216 hanging from the lower surface of the flat plate portion 214.
  • the inside of the tubular portion 216 is a through hole 212 leading to the upper surface of the flat plate portion 214.
  • the portion of the surface 10a of the culture substrate 10 surrounded by each tubular portion 216 is the culture region 12.
  • each culture region 12 is surrounded by the side wall portion 216a of the tubular portion 216, and a plurality of recesses 218 for culturing cells are formed.
  • the arrangement pattern of the recesses 218 in the culture vessel 200 that is, the arrangement pattern of the through holes 212 in the frame body 210 is not particularly limited, and examples thereof include a matrix shape and a staggered shape.
  • the opening shape of the recess 218 in a plan view is not particularly limited, and examples thereof include a round shape and a rectangular shape.
  • the average diameter of the opening of the recess 218, that is, the average diameter of the opening of the through hole 212 is preferably 1.5 mm or more and 40 mm or less, and more preferably 1.7 mm or more and 35 mm or less.
  • the average diameter of the opening of the recess 218 is equal to or more than the lower limit of the above range, it is preferable because it is suitable for drug discovery screening of the formed spheroid.
  • the average diameter of the opening of the recess 218 is equal to or less than the upper limit of the above range, the shaking of the medium is suppressed and the spheroids can be prevented from popping out.
  • the average depth of the recess 218 is preferably 4.0 mm or more and 20 mm or less, and more preferably 5.0 mm or more and 18 mm or less.
  • the average depth of the recesses 218 is equal to or greater than the lower limit of the above range, a sufficient amount of the minimum amount of medium necessary for culturing can be sufficiently added.
  • the average depth of the recess 218 is equal to or less than the upper limit of the above range, the spheroid can be easily taken out in the recess.
  • the number of recesses 218 in the culture vessel 200 is not particularly limited, and can be, for example, 1 or more and 1536 or less.
  • the area of each culture region 12 in the culture container 200 of the microplate shape is preferably 7.0 cm 2 or more 80 cm 2 or less, more preferably 7.5 cm 2 or more 75 cm 2 or less.
  • the area of each culture region 12 can be adjusted by adjusting the diameter of the through hole 212 of the frame body 210.
  • the flask-shaped culture container for example, a culture container in which the culture base material 10 is attached so that the culture region 12 faces upward can be exemplified on the bottom surface of the flask-shaped frame.
  • the method of attaching the culture substrate to the bottom of the frame is not particularly limited, and examples thereof include a method of adhering the culture substrate to the bottom of the frame by pressure bonding, welding, and an adhesive.
  • the adhesive used for adhering the culture substrate is, for example, a liquid or tape-based adhesive using a silicone-based, instantaneous-based, epoxy-based, ultraviolet-curable, polypropylene-based, acrylic-based, rubber-based, or polyethylene-based adhesive. Can be exemplified.
  • the culture vessel is composed of two members, the frame and the culture base, and the amount of warpage of the entire surface of both the culture bases is controlled to 400 ⁇ m or less.
  • the adhesion between the culture substrate and the frame is excellent, and the culture area in the culture vessel is flattened, so that the cells can be easily focused, are less affected by external light, and the cells can be easily observed.
  • a plurality of microwells may be formed on the entire surface 10a of the culture base material 10, and a frame body forming each recess 218. It may be partially formed only in the region surrounded by the side wall portion 216a of 210.
  • one culture substrate was attached to one frame, but the culture container of the present invention is not limited to such a mode.
  • a mode in which a plurality of culture substrates are attached to one frame may be used.
  • the embodiment in which a plurality of culture substrates are attached to one frame is advantageous in that it can be easily applied to mass culture in a large culture vessel such as a large flask shape or a petri dish shape.
  • the culture medium of the present invention is excellent in flatness, even in a mode in which a plurality of culture substrates are attached to one frame, the adhesion is excellent and cells can be easily observed.
  • Example 1 A rectangular plate-shaped base material (average thickness: 1000 ⁇ m) of 150 mm ⁇ 105 mm was molded using polystyrene resin. Next, the entire surface of one surface of the plate-shaped substrate is irradiated with a CO 2 laser to form a plurality of microwells composed of irregularities having continuous curved surfaces as illustrated in FIG. did. After forming the microwells, the base material was sandwiched between a pair of quartz glass flattening plates and subjected to annealing treatment under the conditions of a load of 585 g and a temperature of 80 ° C. for 3 hours to obtain a culture base material.
  • Example 2 A culture substrate was produced in the same manner as in Example 1 except that the annealing treatment was not performed.
  • Adhesion was evaluated based on the presence or absence of medium leakage during cell culture in the evaluation of cell observability. In the visual judgment, the medium with no leakage was judged to have good adhesiveness ( ⁇ ), and the medium with leakage was judged to have poor adhesion ( ⁇ ).
  • Table 1 shows the evaluation of each example.
  • the culture medium of Example 1 in which the amount of warpage of both surfaces is 400 ⁇ m or less is superior to the culture medium of Example 2 in which the amount of warpage exceeds 400 ⁇ m, and has excellent adhesiveness to the frame. Also, cell observation was easy.
  • 10 ... Culture substrate, 10a ... Surface, 12 ... Culture area, 14 ... Microwell, 100 ... Culture container, 110 ... Frame, 112 ... Bottom, 112a ... Bottom, 114 ... Side wall, 200 ... Culture container, 212 ... Through hole, 210 ... frame body, 214 ... flat plate portion, 216 ... tubular portion, 216a ... side wall portion, 218 ... recessed portion.

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Abstract

Le but de la présente invention est de fournir un substrat de culture qui a une excellente adhérence entre un substrat de culture et un corps de cadre et permet une observation facile de cellules ou similaires, et une boîte de culture comprenant le substrat de culture.　L'invention concerne un substrat de culture en forme de plaque (10) dans lequel au moins une partie d'une surface (10a) est une région de culture (12), la région de culture (12) ayant une pluralité de micropuits, et la quantité totale de gauchissement des deux surfaces étant inférieure ou égale à 400 µm. L'invention concerne également une boîte de culture comprenant un substrat de culture (10) et un corps de cadre.
PCT/JP2020/048493 2019-12-27 2020-12-24 Substrat de culture et boîte de culture Ceased WO2021132480A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP3214876U (ja) * 2017-11-30 2018-02-08 Ａｇｃテクノグラス株式会社 培養基材
JP3215918U (ja) * 2017-11-30 2018-04-26 Ａｇｃテクノグラス株式会社 培養基材
WO2018123663A1 (fr) * 2016-12-28 2018-07-05 Ａｇｃテクノグラス株式会社 Substrat de culture cellulaire et son procédé de production
JP2019129748A (ja) * 2018-01-30 2019-08-08 日本ゼオン株式会社 培養容器

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2018123663A1 (fr) * 2016-12-28 2018-07-05 Ａｇｃテクノグラス株式会社 Substrat de culture cellulaire et son procédé de production
JP3214876U (ja) * 2017-11-30 2018-02-08 Ａｇｃテクノグラス株式会社 培養基材
JP3215918U (ja) * 2017-11-30 2018-04-26 Ａｇｃテクノグラス株式会社 培養基材
JP2019129748A (ja) * 2018-01-30 2019-08-08 日本ゼオン株式会社 培養容器

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