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Organoid and three dimensional (3D) cell culture systems are emerging as pivotal models for drug screening, investigating personalized medicine, and understanding human organ development. Organoids are generated from pluripotent stem cells or organ progenitor cells that have differentiated and self-organized into a formation that resembles the structure and function of an intact mammalian organ. Use the table below as a guide to find media, matrix components, growth factors, and small molecules for generating your organoid of interest.
Gasteroids
Stomach epithelial organoids
| Base Media Components | |
|---|---|
| N-2 MAX Supplement | Similar to N-2 |
| N21-MAX Supplement | Similar to B27 Supplement |
| N-Acetylcysteine | |
| Penicillin/Streptomycin | |
| DMEM/F-12 | |
| Glutamine | |
| Organoid Harvesting Solution | |
| Additives for long-term growth | |
|---|---|
| A 83-01 | See all TGF-beta/ALK inhibitors |
| Nicotinamide | |
| SB 202190 | See all p38 MAPK inhibitors |
| Recommended Markers | |
|---|---|
| Pepsinogen C | Chief cells |
| Somatostatin | Enteroendocrine cells |
| TFF1 | Gastric cells |
| TFF2 | Gastric cells |
| MUC5AC | Gastric pit mucous cells |
| Periodic acid-Schiff (PAS) staining | Gastric pit mucous cells |
| MUC6 | Gland mucous cells |
| LGR5 |
Gastric stem cell marker |
| E-Cadherin |
Gastric epithelium |
| References: | |
|---|---|
| Mahe M. et al. (2013) Curr. Protoc. Mouse Biol. 3:217 | |
| Sato T. and H. Clevers (2015) Cell. 161:1700 | |
| Bartfeld S. et al. (2015) Gastroenterology. 148:126 | |
Enteroids
Small intestine epithelial organoids
| Base Media Components | |
|---|---|
| N-2 MAX Supplement | Similar to N-2 |
| N21-MAX Supplement | Similar to B27 Supplement |
| N-Acetylcysteine | |
| Penicillin/Streptomycin | |
| DMEM/F-12 | |
| Glutamine | |
| Organoid Harvesting Solution | |
| Additives for long-term growth | |
|---|---|
| A 83-01 | See all TGF-beta/ALK inhibitors |
| Nicotinamide | |
| SB 202190 | See all p38 MAPK inhibitors |
| Recommended Markers | |
|---|---|
| Alkaline Phosphatase | Enterocytes |
| Villin | Enterocytes |
| Chromogranin A | Enteroendocrine cells |
| Synaptophysin | Enteroendocrine cells |
| MUC2 | Goblet cells |
| Periodic acid-Schiff staining | Goblet cells |
| Lysozyme | Paneth cells |
| LGR5 | Intestinal stem cell marker |
| E-Cadherin | Intestinal epithelium |
| ASCL2/Mash | Crypt cell marker |
Colonoids
Colon epithelial organoids
| Base Media Components | |
|---|---|
| N-2 MAX Supplement | Similar to N-2 |
| N21-MAX Supplement | Similar to B27 Supplement |
| N-Acetylcysteine | |
| Penicillin/Streptomycin | |
| DMEM/F-12 | |
| Glutamine | |
| Organoid Harvesting Solution | |
| Additives for long-term growth | |
|---|---|
| A 83-01 | See all TGF-beta/ALK inhibitors |
| Nicotinamide | |
| SB 202190 | See all p38 MAPK inhibitors |
| Recommended Markers | |
|---|---|
| Ki-67/MKI67 | Cell proliferation marker |
| EphB2 | Colon crypt cell marker |
| Chromogranin A | Enteroendocrine cells |
| EpCAM/TROP-1 | Epithelial cell marker |
| Periodic acid-Schiff staining | Mucous cells |
| LGR5 | Intestinal stem cell marker |
| E-Cadherin | Intestinal epithelium |
| ASCL2/Mash | Crypt cell marker |
Liver Organoids
| Base Media Components | |
|---|---|
| N-2 MAX Supplement | Similar to N-2 |
| N21-MAX Supplement | Similar to B27 Supplement |
| N-Acetylcysteine | |
| Penicillin/Streptomycin | |
| DMEM/F-12 | |
| Glutamine | |
| Organoid Harvesting Solution | |
| Recommended Markers | |
|---|---|
| Cytokeratin 7 | Centrilobular hepatocytes |
| EpCAM/TROP-1 | Epithelial cells |
| CD24 | Hepatocyte progenitors |
| CD44 | Hepatocyte progenitors |
| Glutamine Synthetase | Hepatocyte progenitors |
| SP5 | Hepatocyte transcription factor |
| Cytokeratin 19 | Hepatocytes |
| HNF-1 alpha | Hepatocytes |
| HNF-4 alpha/NR2A1 | Hepatocytes |
| Integrin beta 1/CD29 | Hepatocytes |
| Serum Albumin | Hepatocytes |
| Transthyretin/Prealbumin | Hepatocytes |
| Lgr5/GPR49 | Liver stem cells |
| PROM1 | Liver stem cells |
| References: | |
|---|---|
| Sato T. and H. Clevers (2015) Cell. 161:1700 | |
Pancreatic Organoids
| Base Media Components | |
|---|---|
| N-2 MAX Supplement | Similar to N-2 |
| N21-MAX Supplement | Similar to B27 Supplement |
| N-Acetylcysteine | |
| Penicillin/Streptomycin | |
| DMEM/F-12 | |
| Glutamine | |
| Organoid Harvesting Solution | |
| Recommended Markers | |
|---|---|
| Insulin C-Peptide | Beta cells |
| NeuroD1 | Beta cells |
| NKX6.1 | Endocrine cells |
| HNF-6 | Pancreatic progenitors |
| Neurogenin-3 | Pancreatic progenitors |
| PDX-1 | Pancreatic progenitors |
| SOX9 | Pancreatic progenitors |
| References: | |
|---|---|
| Sato T. and H. Clevers (2015) Cell. 161:1700 | |
| Boj S.F. et al. (2015) Cell. 160:324. | |
Prostate Organoids
| Base Media Components | |
|---|---|
| N-2 MAX Supplement | Similar to N-2 |
| N21-MAX Supplement | Similar to B27 Supplement |
| N-Acetylcysteine | |
| Penicillin/Streptomycin | |
| Glutamine | |
| Organoid Harvesting Solution | |
| Recommended Markers | |
|---|---|
| Cytokeratin 5 | Basal prostate cell |
| p63 | Basal prostate cell |
| Integrin alpha 6/CD49f | Epithelial stem cells |
| TROP-2 | Epithelial stem cells |
| Cytokeratin 8 | Luminal prostate cell |
| Androgen R/NR3C4 | Prostate cells |
| Cytokeratin 18 | Prostate cells |
| TMPRSS2 | Prostate cells |
| NKX3.1 | Prostate-specific transcription factor |
| Laminin gamma 1 | Stromal cells |
| Vimentin | Stromal cells |
| References: | |
|---|---|
| Karthaus W.R. et al. (2014) Cell. 159:163. | |
Kidney Organoids
| Base Media Components | |
|---|---|
| N-2 MAX Supplement | Similar to N-2 |
| N21-MAX Supplement | Similar to B27 Supplement |
| N-Acetylcysteine | |
| Penicillin/Streptomycin | |
| DMEM/F-12 | |
| Glutamine | |
| Holo-Transferrin | |
| Organoid Harvesting Solution | |
| Recommended Markers | |
|---|---|
| Brachyury | Mesoderm |
| LHX-1/LIM1 | Mesoderm |
| Cytokeratin 8 | Uteric bud |
| GATA-3 | Uteric bud |
| GFR alpha-1/GDNFR alpha-1 | Uteric bud |
| HOXB7 | Uteric bud |
| Pax2 | Uteric bud |
| Pax8 | Uteric bud |
| Ret | Uteric bud |
| References: | |
|---|---|
| Xia Y. et al. (2014) Nature Protocols. 9:2693. | |
| Xia Y. et al. (2013) Nature Cell Biology. 15:1507. | |
Brain Organoids
| Base Media Components | |
|---|---|
| N-2 MAX Supplement | Similar to N2 Supplement |
| N21-MAX Supplement | Similar to B27 Supplement |
| N21-MAX Vitamin A Free Supplement | Similar to B27 Vitamin Free Supplement |
| Penicillin/Streptomycin | |
| DMEM/F-12 | |
| GlutaMAX | |
| Insulin | |
| 2-mercaptoethanol | |
| Organoid Harvesting Solution | |
| Recommended Markers | |
|---|---|
| EMX1 | Dorsal cortex marker |
| FOXG1 | Forebrain marker |
| Frizzled-9 | Hippocampus marker |
| Neuropilin-2 | Hippocampus marker |
| Prox1 | Hippocampus marker |
| EOMES | Intermediate progenitor marker |
| Pax6 | Neural induction marker/ventral cortex marker |
| SOX1 | Neural inductions |
| beta-II Tubulin (clone TuJ-1) | Neuronal marker |
| MAP2 | Neuronal marker |
| Vimentin | Neuronal marker |
| References: | |
|---|---|
| Lancaster M.A. and J. A. Knoblich (2014) Nat. Protocols. 9:2329 | |
Lung Organoids
| Base Media Components | |
|---|---|
| N-2 MAX Supplement | Similar to N-2 |
| N21-MAX Supplement | Similar to B27 Supplement |
| N-Acetylcysteine | |
| Penicillin/Streptomycin | |
| DMEM/F-12 | |
| Glutamine | |
| Organoid Harvesting Solution | |
| Recommended Markers | |
|---|---|
| p63 | Basal airway cells |
| ACTTUB | Ciliated cells |
| FoxJ1 | Ciliated cells |
| Uteroglobin/SCGB1A1 | Club cells |
| NKX2.1 | Lung lineage marker |
| ID2 | Lung progenitor marker |
| Lgr5/GPR49 | Lung stem cells |
| alpha-Smooth Muscle Actin | Smooth muscle cells |
| HOPX | Type 1 airway epithelial cells |
| SFTPC | Type 2 airway epithelial cells |
| References: | |
|---|---|
| Dye B.R. et al. (2015) eLIFE. 4:e05098 | |
Inner Ear Organoids
| LIF-2: Base Media Components | |
|---|---|
| N-2 MAX Supplement | Similar to N2 Supplement |
| N21-MAX Supplement | Similar to B27 Supplement |
| Leukemia Inhibitor Factor (LIF) | |
| CHIR 99021 | See all GSK-3 inhibitors |
| PD 0325901 | See all MEK inhibitors |
| DMEM/F-12 | |
| Penicillin/Streptomycin | |
| Glutamine | |
| Organoid Harvesting Solution | |
| Recommended Markers | |
|---|---|
| GATA-3 | Preplacodal ectoderm |
| SIX1 | Preplacodal ectoderm |
| TFAP2A | Preplacodal ectoderm |
| Calretinin | Hair cell marker |
| Myosin 7a | Hair cell marker |
| Pax2 | Hair cell marker |
| SOX2 | Hair cell marker |
| References: | |
|---|---|
| Koehler K.R. and E. Hashino (2014) Nat. Protocols. 9:1229. | |
Cancer Organoids
Cancer organoids should be cultured in similar conditions as to those for which the parent organoid can be grown. In some instances mutations occur in cancer stem cells that allow them to grow in the absence of normal growth factors, such as EGF or FGF. Depending on the cancer organoid it may be possible to grow them in a medium where one or more factors are removed from the normal organoids growth medium.