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Organoid Culture
Organoids are in-vitro 3D cell aggregates derived from stem cells which are not only capable of self-organization and long term self-renewal, but which also exhibit similar function to the tissues from which they were derived.1-4) This is achieved through the use of physical and biochemical cues which are able to recapitulate cells’ natural environment within living tissue. As such, organoids are able to overcome many of the limitations of existing culture models (2D monolayers, 3D aggregates (such as spheroids), animal models, etc.). Current applications for organoid culture systems include those in:
- Developmental Biology5-9)
- Disease Pathology10-22)
- Drug Toxicity / Efficacy Testing23-26)
- Regenerative Medicine27-30)
- Personalized Medicine15,18,26,31,32)
Organoids can be generated by imbedding either primary tissue (human adult stem cells (hAdSCs)) or pluripotent stem cells (induced pluripotent stem cells (iPSCs) or embryonic stem cells (ESCs)) in the appropriate matrix components and applying appropriate signaling molecules / proteins.3)
Some examples of organoid culture conditions can be found below (matrix components, base culture medium, and chemically defined supplements not shown).
| Tissue Type | Origin | Culture Conditions | Reference |
|---|---|---|---|
| Intestine | 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 | |
| Colon | hAdSCs | A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], Y-27623 [Y0018], SB202190, EGF, Rspondin1, Noggin, Wnt-3a | 33 |
| Stomach | 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 | |
| Lung | hAdSCs | A83-01 [A3324], Nicotinamide [N0078], Y-27632 [Y0018], SB202190, Rspondin1, FGF-7, FGF-10, Noggin | 35 |
| hPSCs | hPSC Differentiation: SB431542 [B4003], SANT-2, SU-5402, bFGF, Noggin, SHH, SAG, Activin A Organoid Culture: CHIR 99021 [C2943], SB431542, FGF-4, Noggin | 36 | |
| Brain | hPSCs | Y-27632 [Y0018], Heparin [H0393], 2-Mercaptoethanol [M0058], bFGF, Insulin | 37 |
| Liver | hAdSCs | Organoid Culture: Y-27632 [Y0018], A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], Forskolin [F0855], EGF, Rspondin1, FGF-10, Gastrin, Noggin, Wnt-3a, HGF Hepatocyte Differentiation: 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 | |
| Pancreas | hAdSCs | A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], EGF, Rspondin1, FGF-10, Gastrin, Noggin, Wnt-3a | 20 |
| Kidney | hPSCs | Differentiation: 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 Organoid Culture: CHIR 99021 [C2943], Heparin [H0393], FGF-9, HGF, GDNF | 39, 40 |
| Prostate | 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 | Prostate Differentiation: FGF-10, Activin A, Wnt-10b Organoid Culture: Retinoic Acid [R0064], Testosterone [T0027], EGF, Rspondin1, Noggin | 42 |
References
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- 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
