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オルガノイド培養
オルガノイドとは、試験管内で培養可能な、生体内の組織の特徴を有する幹細胞由来の細胞塊のことです1-4)。物理的な細胞外基質(Extracellular Matrix; ECM)や生化学的なシグナル(増殖因子や阻害剤等)の暴露により自己組織化し、高度な自己複製能を有します。また、生体組織に似た構造だけでなく、それに類似した機能も長期間にわたり維持することができます。そのため、従来の培養モデル(2次元培養法、スフェロイド等他の3次元培養法、動物モデル)の弱点を克服できるとされており、次世代培養法として大いに期待されています。オルガノイド培養が主に応用されている分野は次の通りです。
- 発生生物学5-9)
- 病理学10-22)
- 薬物の有効性・毒性試験23-26)
- 再生医療27-30)
- 個別化医療15,18,26,31,32)
オルガノイドは、検体から採取した初代細胞(ヒト幹細胞 - hAdSCs)または多能性幹細胞(hPSCs – ヒト誘導多能性幹細胞やヒト胚性幹細胞等)を人工細胞外マトリックスの中に包埋し、由来組織に応じたシグナル分子(低分子やタンパク質等)を投与することで樹立されます3)。
以下の表は、由来組織別で典型的なオルガノイド培地成分(マトリックス成分、基本培地、複製成分からなる添加剤は未記載)をリスト化したものです。
由来組織 | 前駆細胞 | 培地成分 | 引用文献 |
---|---|---|---|
小腸 | hAdSCs | A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], Y-27623 [Y0018], SB202190, EGF, Rspondin1, Noggin, Wnt-3a | 33 |
hPSCs | EGF, Rspondin1, FGF-4, Noggin, Wnt-3a, Activin A | 34 | |
大腸 | hAdSCs | A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], Y-27623 [Y0018], SB202190, EGF, Rspondin1, Noggin, Wnt-3a | 33 |
胃 | hAdSCs | A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], Y-27623 [Y0018], EGF, Rspondin, FGF-10, Gastrin, Noggin, Wnt-3a | 11 |
hPSCs | CHIR 99021 [C2943], Y-27623 [Y0018], Retinoic Acid [R0064], EGF, FGF-4, BMP-4, Noggin, Activin A, Wnt-3a | 12 | |
肺 | hAdSCs | A83-01 [A3324], Nicotinamide [N0078], Y-27632 [Y0018], SB202190, Rspondin1, FGF-7, FGF-10, Noggin | 35 |
hPSCs | <分化用> SB431542 [B4003], SANT-2, SU-5402, bFGF, Noggin, SHH, SAG, Activin A <オルガノイド培養用> CHIR 99021 [C2943], SB431542, FGF-4, Noggin | 36 | |
脳 | hPSCs | Y-27632 [Y0018], Heparin [H0393], 2-Mercaptoethanol [M0058], bFGF, Insulin | 37 |
肝臓 | hAdSCs | <オルガノイド培養用> Y-27632 [Y0018], A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], Forskolin [F0855], EGF, Rspondin1, FGF-10, Gastrin, Noggin, Wnt-3a, HGF <オルガノイド樹立後分化用> A83-01 [A3324], DAPT [D4257], Dexamethasone [D1961], EGF, FGF-19, BMP-7, Gastrin, HGF | 17 |
hPSCs | bFGF, BMP-4, Activin A, HGF, Oncostatin M | 38 | |
膵臓 | hAdSCs | A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], EGF, Rspondin1, FGF-10, Gastrin, Noggin, Wnt-3a | 20 |
腎臓 | hPSCs | <分化用> CHIR 99021 [C2943], 2-Mercaptoethanol [M0058], Heparin [H0393], Retinoic Acid [R0064], 1-Thioglycerol [T0905], bFGF, FGF-9, BMP-2, BMP-4, Insulin, Activin A, Holo-transferrin <オルガノイド培養用> CHIR 99021 [C2943], Heparin [H0393], FGF-9, HGF, GDNF | 39, 40 |
前立腺 | hAdSCs | Y-27623 [Y0018], A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], SB202190, Prostaglandin E2 [P1884], Testosterone [T0027], EGF, Rspondin1, FGF-10, bFGF, Noggin | 41 |
hPSCs | <分化用> FGF-10, Activin A, Wnt-10b <オルガノイド培養用> Retinoic Acid [R0064], Testosterone [T0027], EGF, Rspondin1, Noggin | 42 |
引用文献
=======================>>[引用文献をご覧になる場合はこちらをクリック]<<=======================
- 1) Cytosystems dynamics in self-organization of tissue architecture
- 2) Organogenesis in a dish: Modeling development and disease using organoid technologies
- 3) Human organoids: model systems for human biology and medicine
- 4) Organoid and Spheroid Tumor Models: Techniques and Applications
- 5) Modeling Development and Disease with Organoids
- 6) Modeling mouse and human development using organoid cultures
- 7) Organoid Models of Human Gastrointestinal Development and Disease
- 8) Self-organizing optic-cup morphogenesis in three-dimensional culture
- 9) Multilineage communication regulates human liver bud development from pluripotency
- 10) Life-threatening influenza and impaired interferon amplification in human IRF7 deficiency
- 11) In Vitro Expansion of Human Gastric Epithelial Stem Cells and Their Responses to Bacterial Infection
- 12) Modelling human development and disease in pluripotent stem-cell-derived gastric organoids
- 13) Recapitulation of hepatitis B virus–host interactions in liver organoids from human induced pluripotent stem cells
- 14) Cerebral organoids model human brain development and microcephaly
- 15) Directed differentiation of human pluripotent stem cells into mature airway epithelia expressing functional CFTR protein
- 16) Functional Repair of CFTR by CRISPR/Cas9 in Intestinal Stem Cell Organoids of Cystic Fibrosis Patients
- 17) Long-Term Culture of Genome-Stable Bipotent Stem Cells from Adult Human Liver
- 18) Prospective Derivation of a Living Organoid Biobank of Colorectal Cancer Patients
- 19) Organoid Cultures Derived from Patients with Advanced Prostate Cancer
- 20) Organoid Models of Human and Mouse Ductal Pancreatic Cancer
- 21) Human primary liver cancer–derived organoid cultures for disease modeling and drug screening
- 22) A Living Biobank of Breast Cancer Organoids Captures Disease Heterogeneity
- 23) Kidney organoids from human iPS cells contain multiple lineages and model human nephrogenesis
- 24) Reverse engineering liver buds through self-driven condensation and organization towards medical application
- 25) Synergistic Engineering: Organoids Meet Organs-on-a-Chip
- 26) Patient-derived organoids model treatment response of metastatic gastrointestinal cancers
- 27) Vascularized and Complex Organ Buds from Diverse Tissues via Mesenchymal Cell-Driven Condensation
- 28) Vascularized and functional human liver from an iPSC-derived organ bud transplant
- 29) Functional engraftment of colon epithelium expanded in vitro from a single adult Lgr5+ stem cell
- 30) In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration
- 31) Preserved genetic diversity in organoids cultured from biopsies of human colorectal cancer metastases
- 32) Molecular dissection of colorectal cancer in pre-clinical models identifies biomarkers predicting sensitivity to EGFR inhibitors
- 33) Long-term Expansion of Epithelial Organoids From Human Colon, Adenoma, Adenocarcinoma, and Barrett’s Epithelium
- 34) Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro
- 35) Long‐term expanding human airway organoids for disease modeling
- 36) In vitro generation of human pluripotent stem cell derived lung organoids
- 37) Generation of cerebral organoids from human pluripotent stem cells
- 38) Highly efficient generation of human hepatocyte-like cells from induced pluripotent stem cells
- 39) Generation of kidney organoids from human pluripotent stem cells
- 40) Engineered Kidney Tubules for Modeling Patient-Specific Diseases and Drug Discovery
- 41) Organoid culture systems for prostate epithelial and cancer tissue
- 42) Directed Differentiation of Human Embryonic Stem Cells into Prostate Organoids In Vitro and its Perturbation by Low-Dose Bisphenol A Exposure