JP7107595B2 - 多能性幹細胞由来結膜細胞の誘導方法 - Google Patents
多能性幹細胞由来結膜細胞の誘導方法 Download PDFInfo
- Publication number
- JP7107595B2 JP7107595B2 JP2020520350A JP2020520350A JP7107595B2 JP 7107595 B2 JP7107595 B2 JP 7107595B2 JP 2020520350 A JP2020520350 A JP 2020520350A JP 2020520350 A JP2020520350 A JP 2020520350A JP 7107595 B2 JP7107595 B2 JP 7107595B2
- Authority
- JP
- Japan
- Prior art keywords
- cells
- conjunctival
- conjunctival epithelial
- stem
- cell
- Prior art date
- Legal status (The legal status 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 status listed.)
- Active
Links
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
- G01N33/5064—Endothelial cells
-
- 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
- C12N5/0618—Cells of the nervous system
- C12N5/0621—Eye cells, e.g. cornea, iris pigmented cells
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
- G01N33/5058—Neurological cells
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
- G01N33/5073—Stem cells
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/11—Epidermal growth factor [EGF]
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/115—Basic fibroblast growth factor (bFGF, FGF-2)
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/10—Growth factors
- C12N2501/117—Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12)
-
- 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
- C12N2501/00—Active agents used in cell culture processes, e.g. differentation
- C12N2501/70—Enzymes
- C12N2501/72—Transferases (EC 2.)
- C12N2501/727—Kinases (EC 2.7.)
-
- 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
- C12N2503/00—Use of cells in diagnostics
- C12N2503/02—Drug screening
-
- 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
- C12N2506/00—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells
- C12N2506/45—Differentiation of animal cells from one lineage to another; Differentiation of pluripotent cells from artificially induced pluripotent stem cells
-
- 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
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/50—Proteins
- C12N2533/52—Fibronectin; Laminin
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- Cell Biology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- General Engineering & Computer Science (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Ophthalmology & Optometry (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Toxicology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Developmental Biology & Embryology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Materials For Medical Uses (AREA)
Description
〔1〕 多能性幹細胞のコロニーをEGF(Epidermal Growth Factor)シグナル伝達活性化因子を含む培地にて培養して分化誘導させることを特徴とする、結膜上皮細胞、結膜杯細胞ならびに結膜上皮幹・前駆細胞の誘導方法。
〔2〕 多能性幹細胞のコロニーをEGFシグナル伝達活性化因子を含む培地にて培養し、SEAM細胞集団(self-formed ectodermal autonomous multi-zone細胞集団)を形成させる、前記〔1〕記載の誘導方法。
〔3〕 多能性幹細胞のコロニーをEGF(Epidermal Growth Factor)シグナル伝達活性化因子を含む培地にて培養してSEAM細胞集団(self-formed ectodermal autonomous multi-zone細胞集団)を形成させ、前記SEAM細胞集団から、ITGβ4、SSEA-4及びCD200の抗体の発現程度を指標にして細胞を単離することを特徴とする、結膜上皮細胞、結膜杯細胞ならびに結膜上皮幹・前駆細胞の製造方法。
〔4〕 単離した細胞がCD200陰性である、前記〔3〕記載の製造方法。
〔5〕 単離した細胞がSSEA-4弱陽性/ITGβ4陽性細胞である、前記〔3〕又は〔4〕記載の製造方法。
〔6〕 単離された細胞が結膜上皮幹・前駆細胞である、前記〔5〕記載の製造方法。
〔7〕 単離した細胞がSSEA-4弱陽性/ITGβ4陰性細胞及びSSEA-4陽性/ITGβ4陰性細胞から選ばれる、前記〔3〕又は〔4〕記載の製造方法。
〔8〕 単離された細胞が結膜上皮細胞及び/又は結膜杯細胞である、前記〔7〕記載の製造方法。
〔9〕 前記〔1〕~〔6〕いずれか記載の方法により得られた結膜上皮幹・前駆細胞をKGF(Keratinocyte Growth Factor)を含む培地にて培養する工程を含む、結膜上皮細胞シートの製造方法。
〔10〕 前記〔1〕~〔6〕いずれか記載の方法により得られた結膜上皮幹・前駆細胞をFGFR2b(Fibroblast Growth Factor Receptor 2b)活性化因子を含む培地にて培養する工程を含む、結膜上皮細胞シートの製造方法。
〔11〕 前記〔1〕~〔8〕いずれか記載の方法により得られた結膜上皮細胞、結膜杯細胞及び/又は結膜上皮幹・前駆細胞を培養する工程を含む、結膜に関連する疾患の薬剤スクリーニング方法。
iPS細胞からの分化誘導には、self-formed ectodermal autonomous multi-zone(SEAM)法を用いた(Hayashi et al. Nature. 2016 Mar 17;531(7594):376-80., Hayashi et al., Nature Protocols, 2017, 12(4), 683-696, doi: 10.1038/nprot.2017.007)。
実施例1で得られたSEAM細胞集団から、セルソーターSH800(SONY)にてソーティングしてCD200陰性細胞を抽出後、SSEA-4、ITGβ4にて展開し、図2に示す6つの画分に分画した。各画分の定義は、SSEA4-陰性/ITGβ4陽性細胞(P1画分)、SSEA-4弱陽性/ITGβ4陽性細胞(P2画分)、SSEA-4陽性/ITGβ4陽性細胞(P3画分)、SSEA-4陰性/ITGβ4陰性細胞(P4画分)、SSEA-4弱陽性/ITGβ4陰性細胞(P5画分)、SSEA-4陽性/ITGβ4陰性細胞(P6画分)の通りである。
実施例2で得られた6つの画分それぞれを、QIAzol Lysis Reagent(QIAGEN)を用いて回収し、RT-qPCRに供し、各種マーカー発現を評価した。結果を図3に示す。なお、図中、P4画分由来細胞(n=3)、その他の画分由来細胞(n=5)の結果であり、エラーバーは標準誤差を示した。
実施例2で抽出されたP5・6画分由来細胞を、BD Cytofix/Cytoperm(登録商標)Kit(554714、BD Biosciences)にて固定、膜透過処理後、DMEM/F12培地に50,000cells/mLとなるよう懸濁し、200μL/カラムとして、サイトスピン(A78300003、Thermo Fisher Scientific)にてスライドに細胞標本を作製した。続いて、5%NST(5% Normal Donkey Serum, 0.3% Tritonを含むTBS;T903、TaKaRa Bio)でブロッキング(室温、1時間)後に、抗MUC5AC抗体(1:200;sc-21701、Santa Cruz Biotechnology)、抗K13抗体(1:200;ab16112、abcam)、抗K12抗体(1:200;sc-17098、Santa Cruz Biotechnology)、抗PAX6抗体(1:300;PRB-278P、COVANCE)を用いて1次抗体反応(4℃、overnight)を行った。TBSにて5分間、2回洗浄後、Alexa Fluor 488、Alexa Fluor 568またはAlexa Fluor 647で標識した2次抗体(1:200;Life Technologies)及びHoechst33342(1:100;H3570、Molecular Probes)で処理(室温、1時間)後、蛍光顕微鏡で観察した。結果を図4に示す。
実施例2と同様にして得られたP2画分由来細胞を、ラミニン511E8フラグメント(0.5μg/cm2)でコーティングした細胞培養用インサート上で2~3週間培養した。培養には、KGF培地〔10ng/mL KGF(Wako)、10μM Y-27632及び2% B27 Supplement、及び1% penicillin-streptomycin solutionを含有させたDMEM/F-12〕またはEGF培地〔10ng/mL EGF、10μM Y-27632及び2% B27 Supplement、及び1% penicillin-streptomycin solutionを含有させたDMEM/F-12〕を使用した。
実施例1において、結膜用分化培地におけるEGFを、transforming growth factorα(TGFα)又はamphi-regulin(AR)に変更する以外は、実施例1と同様にして分化誘導を行なった。その後、実施例2と同様にしてP2画分由来細胞を抽出後、実施例5と同様にしてKGF培地で培養し、得られた細胞シートの染色を行なってMUC5ACの発現を確認した。
実施例5において、結膜上皮幹・前駆細胞の培養培地におけるKGFを、FGFR2b活性化因子(FGF10)に変更する以外は、実施例5と同様にして培養し、得られた細胞シートを実施例3と同様にしてRT-qPCRに供して、MUC5ACの発現量を評価した。
Claims (11)
- 多能性幹細胞のコロニーをEGF(Epidermal Growth Factor)シグナル伝達活性化因子を含む培地にて培養して分化誘導させることを特徴とする、結膜上皮細胞、結膜杯細胞ならびに結膜上皮幹・前駆細胞の誘導方法。
- 多能性幹細胞のコロニーをEGFシグナル伝達活性化因子を含む培地にて培養し、SEAM細胞集団(self-formed ectodermal autonomous multi-zone細胞集団)を形成させる、請求項1記載の誘導方法。
- 多能性幹細胞のコロニーをEGF(Epidermal Growth Factor)シグナル伝達活性化因子を含む培地にて培養してSEAM細胞集団(self-formed ectodermal autonomous multi-zone細胞集団)を形成させ、前記SEAM細胞集団から、ITGβ4、SSEA-4及びCD200の発現程度を指標にして細胞を単離することを特徴とする、結膜上皮細胞、結膜杯細胞ならびに結膜上皮幹・前駆細胞の製造方法。
- 単離した細胞がCD200陰性である、請求項3記載の製造方法。
- 単離した細胞がSSEA-4弱陽性/ITGβ4陽性細胞である、請求項3又は4記載の製造方法。
- 単離された細胞が結膜上皮幹・前駆細胞である、請求項5記載の製造方法。
- 単離した細胞がSSEA-4弱陽性/ITGβ4陰性細胞及びSSEA-4陽性/ITGβ4陰性細胞から選ばれる、請求項3又は4記載の製造方法。
- 単離された細胞が結膜上皮細胞及び/又は結膜杯細胞である、請求項7記載の製造方法。
- 請求項3~6いずれか記載の方法により結膜上皮幹・前駆細胞を製造する工程、及び得られた結膜上皮幹・前駆細胞をKGF(Keratinocyte Growth Factor)を含む培地にて培養する工程を含む、結膜上皮細胞シートの製造方法。
- 請求項3~6いずれか記載の方法により結膜上皮幹・前駆細胞を製造する工程、及び得られた結膜上皮幹・前駆細胞をFGFR2b(Fibroblast Growth Factor Receptor 2b)活性化因子を含む培地にて培養する工程を含む、結膜上皮細胞シートの製造方法。
- 請求項3~8いずれか記載の方法により結膜上皮細胞、結膜杯細胞及び/又は結膜上皮幹・前駆細胞を製造する工程、及び得られた結膜上皮細胞、結膜杯細胞及び/又は結膜上皮幹・前駆細胞を培養する工程を含む、結膜に関連する疾患の薬剤スクリーニング方法。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2018098954 | 2018-05-23 | ||
JP2018098954 | 2018-05-23 | ||
PCT/JP2019/020364 WO2019225672A1 (ja) | 2018-05-23 | 2019-05-22 | 多能性幹細胞由来結膜細胞の誘導方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
JPWO2019225672A1 JPWO2019225672A1 (ja) | 2021-05-13 |
JP7107595B2 true JP7107595B2 (ja) | 2022-07-27 |
Family
ID=68616983
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2020520350A Active JP7107595B2 (ja) | 2018-05-23 | 2019-05-22 | 多能性幹細胞由来結膜細胞の誘導方法 |
Country Status (8)
Country | Link |
---|---|
US (1) | US20210207087A1 (ja) |
EP (1) | EP3783097A4 (ja) |
JP (1) | JP7107595B2 (ja) |
KR (1) | KR20210015789A (ja) |
CN (1) | CN112189049B (ja) |
AU (1) | AU2019273567A1 (ja) |
SG (1) | SG11202011365XA (ja) |
WO (1) | WO2019225672A1 (ja) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023190745A1 (ja) * | 2022-04-01 | 2023-10-05 | 国立大学法人大阪大学 | 結膜上皮マーカーおよびその利用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2006129724A (ja) | 2004-11-02 | 2006-05-25 | Senju Pharmaceut Co Ltd | ラクリチン活性を有する化合物のスクリーニング方法 |
WO2009011139A1 (ja) | 2007-07-13 | 2009-01-22 | Mitsubishi Tanabe Pharma Corporation | 細胞の単離方法、細胞用無血清培養培地および細胞の培養方法 |
WO2016114285A1 (ja) | 2015-01-15 | 2016-07-21 | 国立大学法人大阪大学 | 多能性幹細胞からの角膜上皮細胞の分化誘導 |
-
2019
- 2019-05-22 AU AU2019273567A patent/AU2019273567A1/en active Pending
- 2019-05-22 KR KR1020207032930A patent/KR20210015789A/ko active Search and Examination
- 2019-05-22 EP EP19807067.4A patent/EP3783097A4/en active Pending
- 2019-05-22 SG SG11202011365XA patent/SG11202011365XA/en unknown
- 2019-05-22 CN CN201980034709.4A patent/CN112189049B/zh active Active
- 2019-05-22 US US17/057,021 patent/US20210207087A1/en active Pending
- 2019-05-22 WO PCT/JP2019/020364 patent/WO2019225672A1/ja active Application Filing
- 2019-05-22 JP JP2020520350A patent/JP7107595B2/ja active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2006129724A (ja) | 2004-11-02 | 2006-05-25 | Senju Pharmaceut Co Ltd | ラクリチン活性を有する化合物のスクリーニング方法 |
WO2009011139A1 (ja) | 2007-07-13 | 2009-01-22 | Mitsubishi Tanabe Pharma Corporation | 細胞の単離方法、細胞用無血清培養培地および細胞の培養方法 |
WO2016114285A1 (ja) | 2015-01-15 | 2016-07-21 | 国立大学法人大阪大学 | 多能性幹細胞からの角膜上皮細胞の分化誘導 |
Non-Patent Citations (3)
Title |
---|
Nature Protocols,2017年,Vol.12, No.4,pp.683-696 |
Nature,2016年,Vol.531,pp.376-380 |
PLoS ONE,2011年,Vol.6, No.9, e24194,pp.1-13 |
Also Published As
Publication number | Publication date |
---|---|
CN112189049A (zh) | 2021-01-05 |
AU2019273567A1 (en) | 2020-12-03 |
SG11202011365XA (en) | 2020-12-30 |
EP3783097A4 (en) | 2021-04-14 |
KR20210015789A (ko) | 2021-02-10 |
JPWO2019225672A1 (ja) | 2021-05-13 |
WO2019225672A1 (ja) | 2019-11-28 |
CN112189049B (zh) | 2023-12-15 |
US20210207087A1 (en) | 2021-07-08 |
EP3783097A1 (en) | 2021-02-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2014303330B2 (en) | Method for producing dopaminergic neurons | |
JP6756610B2 (ja) | 多分化能細胞および多能性細胞の分化を方向付けることによって発生させる皮質介在ニューロンおよびその他のニューロン細胞 | |
EP3246394B1 (en) | Method for inducing differentiation of corneal epithelial cells from pluripotent stem cells | |
JP2012523240A (ja) | 幹細胞培養のための方法および組成物 | |
US20200283735A1 (en) | Organoid and method for producing the same | |
JP2016534738A (ja) | 心外膜細胞を形成するための方法及び組成物 | |
JPWO2007010858A1 (ja) | 骨格筋組織由来の単一細胞よりクローン化した多能性幹細胞 | |
KR102208889B1 (ko) | 다능성줄기세포의 분화 제어 방법 | |
KR100677054B1 (ko) | 제대혈로부터 다분화능 전구세포를 분리하여 배양하는 방법 및 이의 분화 유도방법 | |
JP7107595B2 (ja) | 多能性幹細胞由来結膜細胞の誘導方法 | |
JP7016185B2 (ja) | 幹細胞由来涙腺組織の作製方法 | |
US20220313733A1 (en) | Method for producing renal interstitial cell | |
US20220010271A1 (en) | Method for producing brain organoids | |
WO2021187601A1 (ja) | 心筋細胞の精製方法 | |
KR20080094431A (ko) | 말초 혈액 유래 단핵 세포로부터 신경 전구 세포를 분리,배양 및 분화하는 방법 | |
CA3224178A1 (en) | Method for producing cerebral cortical cell preparation derived from human pluripotent stem cells | |
AU2022235278A1 (en) | Methods of generating oligodendrocyte progenitor cells and use thereof |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20201020 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20211012 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20211206 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20220222 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20220224 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20220628 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20220707 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 7107595 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |