JP2015070832A - 細胞培養用製品及びその製造方法 - Google Patents
細胞培養用製品及びその製造方法 Download PDFInfo
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- hydrophilic copolymer
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- 238000004113 cell culture Methods 0.000 title claims abstract description 82
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 23
- 229920001480 hydrophilic copolymer Polymers 0.000 claims abstract description 168
- 239000004372 Polyvinyl alcohol Substances 0.000 claims abstract description 87
- 229920002451 polyvinyl alcohol Polymers 0.000 claims abstract description 87
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 85
- 125000002843 carboxylic acid group Chemical group 0.000 claims abstract description 58
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 45
- 125000003158 alcohol group Chemical group 0.000 claims abstract description 42
- 239000000758 substrate Substances 0.000 claims abstract description 41
- 238000000034 method Methods 0.000 claims abstract description 20
- 238000004132 cross linking Methods 0.000 claims description 48
- 102100037362 Fibronectin Human genes 0.000 claims description 25
- 108010067306 Fibronectins Proteins 0.000 claims description 25
- 102100035140 Vitronectin Human genes 0.000 claims description 25
- 108010031318 Vitronectin Proteins 0.000 claims description 25
- 238000006243 chemical reaction Methods 0.000 claims description 24
- 239000012634 fragment Substances 0.000 claims description 24
- 230000035484 reaction time Effects 0.000 claims description 23
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 22
- 102000015636 Oligopeptides Human genes 0.000 claims description 20
- 108010038807 Oligopeptides Proteins 0.000 claims description 20
- 102000004169 proteins and genes Human genes 0.000 claims description 20
- 108090000623 proteins and genes Proteins 0.000 claims description 20
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 12
- 230000008569 process Effects 0.000 claims description 12
- JAHNSTQSQJOJLO-UHFFFAOYSA-N 2-(3-fluorophenyl)-1h-imidazole Chemical compound FC1=CC=CC(C=2NC=CN=2)=C1 JAHNSTQSQJOJLO-UHFFFAOYSA-N 0.000 claims description 11
- LVHBHZANLOWSRM-UHFFFAOYSA-N methylenebutanedioic acid Natural products OC(=O)CC(=C)C(O)=O LVHBHZANLOWSRM-UHFFFAOYSA-N 0.000 claims description 11
- 229920002432 poly(vinyl methyl ether) polymer Polymers 0.000 claims description 11
- 229920002689 polyvinyl acetate Polymers 0.000 claims description 11
- 239000011118 polyvinyl acetate Substances 0.000 claims description 11
- 229920002554 vinyl polymer Polymers 0.000 claims description 11
- 150000001413 amino acids Chemical class 0.000 claims description 10
- 229920001184 polypeptide Polymers 0.000 claims description 10
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 10
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 claims description 6
- 239000002202 Polyethylene glycol Substances 0.000 claims 1
- 125000003827 glycol group Chemical group 0.000 claims 1
- 229920001223 polyethylene glycol Polymers 0.000 claims 1
- 210000000130 stem cell Anatomy 0.000 abstract description 46
- 230000024245 cell differentiation Effects 0.000 abstract description 16
- 238000012258 culturing Methods 0.000 abstract description 7
- 230000001105 regulatory effect Effects 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 32
- 125000003277 amino group Chemical group 0.000 description 15
- 230000001276 controlling effect Effects 0.000 description 14
- 239000006143 cell culture medium Substances 0.000 description 12
- 210000001671 embryonic stem cell Anatomy 0.000 description 12
- 239000000126 substance Substances 0.000 description 12
- 230000004069 differentiation Effects 0.000 description 11
- 238000007334 copolymerization reaction Methods 0.000 description 9
- 238000010586 diagram Methods 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- 230000000052 comparative effect Effects 0.000 description 7
- 210000004263 induced pluripotent stem cell Anatomy 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 229920000193 polymethacrylate Polymers 0.000 description 7
- 210000004504 adult stem cell Anatomy 0.000 description 6
- 230000010261 cell growth Effects 0.000 description 6
- 239000003102 growth factor Substances 0.000 description 6
- 108020003175 receptors Proteins 0.000 description 5
- 102000005962 receptors Human genes 0.000 description 5
- 102000009618 Transforming Growth Factors Human genes 0.000 description 4
- 108010009583 Transforming Growth Factors Proteins 0.000 description 4
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 4
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 4
- 108010044426 integrins Proteins 0.000 description 4
- 102000006495 integrins Human genes 0.000 description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 4
- 229920001577 copolymer Polymers 0.000 description 3
- 210000000663 muscle cell Anatomy 0.000 description 3
- 210000000963 osteoblast Anatomy 0.000 description 3
- 230000009471 action Effects 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 108010030465 Integrin alpha6beta1 Proteins 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- 125000004036 acetal group Chemical group 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- ZHNUHDYFZUAESO-UHFFFAOYSA-N formamide Substances NC=O ZHNUHDYFZUAESO-UHFFFAOYSA-N 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000003061 neural cell Anatomy 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0696—Artificially induced pluripotent stem cells, e.g. iPS
-
- 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
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/0062—General methods for three-dimensional culture
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0603—Embryonic cells ; Embryoid bodies
- C12N5/0606—Pluripotent embryonic cells, e.g. embryonic stem cells [ES]
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0625—Epidermal cells, skin cells; Cells of the oral mucosa
- C12N5/0627—Hair cells
- C12N5/0628—Hair stem cells; Hair progenitors
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/30—Synthetic polymers
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/50—Proteins
- C12N2533/52—Fibronectin; Laminin
Abstract
【解決手段】この細胞培養用製品は、基板と、親水性共重合体層と、複数のペプチド鎖とを具備する。基板は、表面を有し、親水性共重合体層は、基材の表面に設けられる。親水性共重合体層は、複数のポリビニルアルコール単位、複数のポリビニルアルコール誘導体単位及び複数のカルボン酸基含有単位によって共重合されたものである。ペプチド鎖は、それぞれ親水性の共重合体層の表面に結合される。細胞培養用製品の製造方法、未分化幹細胞を培養する方法、及び幹細胞分化を調整・制御する方法も、ここで開示されている。
【選択図】図1A
Description
110、210 基板
111、121、125、127、211、221 表面
120a、120b、120c、220a、220b、220c 親水性共重合体層
122 第1弾性層
124 第2弾性層
126 第3弾性層
130、230 ペプチド鎖
310 架橋反応液
410、420、430、440、510、520、530、610、620、630、640 工程
710 細胞コロニー
720 細胞
721 受容体
D1、D2 厚さ
Claims (23)
- 表面を有する基板、
前記基板の前記表面に設けられ、複数のポリビニルアルコール単位、複数のポリビニルアルコール誘導体単位及び複数のカルボン酸基含有単位によって共重合された親水性共重合体層、並びに
前記親水性共重合体層の表面にそれぞれに結合される複数のペプチド鎖、
を具備する細胞培養用製品。 - ポリビニルアルコール誘導体単位が、ポリビニルアセテート、ポリビニルメチルエーテル又はポリビニルエチルエーテルを含む、請求項1に記載の細胞培養用製品。
- カルボン酸基含有単位が、イタコン酸を含む、請求項1または2に記載の細胞培養用製品。
- 親水性共重合体層において、前記ポリビニルアルコール単位、前記ポリビニルアルコール誘導体単位及び前記カルボン酸基含有単位の含有量が、それぞれ約65〜98.7%、0〜30%および1.3〜5%である、請求項1〜3のいずれか一項に記載の細胞培養用製品。
- 親水性共重合体層において、前記ポリビニルアルコール単位、前記ポリビニルアルコール誘導体単位及び前記カルボン酸基含有単位の含有量が、それぞれ約85%、10%および5%である、請求項1〜4のいずれか一項に記載の細胞培養用製品。
- 親水性共重合体層が、弾性係数を有する、請求項1〜5のいずれか一項に記載の細胞培養用製品。
- 弾性係数が、約1kPa〜100kPaの範囲内にある、請求項6に記載の細胞培養用製品。
- ペプチド鎖が、オリゴペプチド、ポリペプチド、タンパク質又はこれらの組み合わせを含む、請求項1〜7のいずれか一項に記載の細胞培養用製品。
- オリゴペプチドが、フィブロネクチンのCS‐1断片又はビトロネクチンの断片を含む、請求項8に記載の細胞培養用製品。
- タンパク質が、フィブロネクチン又はビトロネクチンを含む、請求項8に記載の細胞培養用製品。
- ペプチド鎖のアミノ酸が、アミド結合によって、カルボン酸基含有単位にそれぞれに結合される、請求項1〜10のいずれか一項に記載の細胞培養用製品。
- 表面を有する基板を提供する工程と、
共重合反応によって、複数のポリビニルアルコール単位、複数のポリビニルアルコール誘導体単位及び複数のカルボン酸基含有単位を共重合させた親水性共重合体層を、前記基板に形成する工程と、
前記親水性共重合体層における前記ポリエチレングリコール単位同士を架橋させる架橋反応を行う工程と、
複数のペプチド鎖をそれぞれに前記親水性共重合体層の表面に結合する工程
とを含む、細胞培養用製品の製造方法。 - 親水性共重合体層を形成する工程が、親水性共重合体溶液を調製し、前記親水性共重合体溶液を前記基板の前記表面に塗布することを含む、請求項12に記載の製造方法。
- 親水性共重合体溶液が、約0.05〜0.2wt%の濃度で親水性共重合体を含む、請求項13に記載の製造方法。
- ポリビニルアルコール誘導体単位が、ポリビニルアセテート、ポリビニルメチルエーテル又はポリビニルエチルエーテルを含む、請求項12〜14のいずれか一項に記載の製造方法。
- カルボン酸基含有単位が、イタコン酸を含む、請求項12〜15のいずれか一項に記載の製造方法。
- ペプチド鎖が、オリゴペプチド、ポリペプチド、タンパク質又はこれらの組み合わせを含む、請求項12〜16のいずれか一項に記載の製造方法。
- オリゴペプチドが、フィブロネクチンの断片(CS‐1)又はビトロネクチンの断片を含む、請求項17に記載の製造方法。
- タンパク質が、フィブロネクチン又はビトロネクチンを含む、請求項17に記載の製造方法。
- 架橋反応を行う工程が、硫酸の存在下で、親水性共重合体層におけるポリビニルアルコール単位をグルタルアルデヒドで架橋することを含む、請求項12〜19のいずれか一項に記載の製造方法。
- 架橋反応を行う工程が、前記架橋反応の架橋反応時間を制御することにより、親水性共重合体層の弾性係数を調整・制御する工程を含む、請求項12〜20のいずれか一項に記載の製造方法。
- 架橋反応時間が、約0.5時間〜48時間である、請求項21に記載の製造方法。
- 弾性係数が、約1kPa〜100kPaの範囲内にある、請求項21または22に記載の製造方法。
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JPWO2020230885A1 (ja) * | 2019-05-15 | 2020-11-19 | ||
JP2022079559A (ja) * | 2015-04-22 | 2022-05-26 | バークレー ライツ,インコーポレイテッド | マイクロ流体細胞培養 |
US11801508B2 (en) | 2016-05-26 | 2023-10-31 | Berkeley Lights, Inc. | Covalently modified surfaces, kits, and methods of preparation and use |
US11964275B2 (en) | 2015-10-27 | 2024-04-23 | Berkeley Lights, Inc. | Microfluidic apparatus having an optimized electrowetting surface and related systems and methods |
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TWI601817B (zh) * | 2013-10-02 | 2017-10-11 | 國立中央大學 | 細胞培養製品及其製造方法 |
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Also Published As
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TWI601817B (zh) | 2017-10-11 |
TW201514298A (zh) | 2015-04-16 |
US9902941B2 (en) | 2018-02-27 |
US20150093828A1 (en) | 2015-04-02 |
US10336986B2 (en) | 2019-07-02 |
JP5944931B2 (ja) | 2016-07-05 |
US20180230437A1 (en) | 2018-08-16 |
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