Organoid and 3D Culture Reagents

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.

View GMP-grade proteins for translational research.

 

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  
3D Growth Matrix Components
Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 See all matrix products
EGF  
FGF-10 See all FGF Ligands
Noggin See all TGF-beta modulators
R-Spondin 1 See all R-Spondins
Wnt-3a See all Wnts
Gastrin  
Y-27623 See more ROCK inhibitors
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  
3D Growth Matrix Components
Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 See all matrix products
EGF  
Noggin See all TGF-beta modulators
R-Spondin 1 See all R-Spondins
Wnt-3a See all Wnts
A 83-01 See all TGF-beta/ALK inhibitor
SB 202190 See all p38 MAPK inhibitors
Y-27623 See all ROCK inhibitors
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
References:
Mahe M. et al. (2013) Curr. Protoc. Mouse Biol. 3:217
Sato T. et al. (2011) Gastroenterology. 141:1762
Sato T. and H. Clevers (2015) Cell. 161:1700
Sato T. and H. Clevers (2009) Nature. 459:262

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  
3D Growth Matrix Components
Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 See all matrix products
EGF  
Noggin See all TGF-beta modulators
R-Spondin 1 See all R-Spondins
Wnt-3a See all Wnts
A 83-01 See all TGF-beta/ALK inhibitor
SB 202190 See all p38 MAPK inhibitors
Y-27623 See all ROCK inhibitors
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
References:
Mahe M. et al. (2013) Curr. Protoc. Mouse Biol. 3:217
Sato T. et al. (2011) Gastroenterology. 141:1762
Sato T. and H. Clevers (2015) Cell. 161:1700
Jung P. et al. (2011) Nature Medicine. 17:1225

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  
3D Growth Matrix Components
Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 See all matrix products
EGF  
FGF-10 See all FGF Ligands
HGF  
Noggin See all TGF-beta modulators
R-Spondin 1 See all R-Spondins
Wnt-3a See all Wnts
A 83-01 See all TGF-beta/ALK inhibitor
DAPT See all Gamma-secretase inhibitors
Forskolin  
Nicotinamide  
Prostaglandin E2  
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  
3D Growth Matrix Components
Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 See all matrix products
EGF  
FGF-10 See all FGF Ligands
Noggin See all TGF-beta modulators
R-Spondin 1 See all R-Spondins
Wnt-3a See all Wnts
A 83-01 See all TGF-beta/ALK inhibitor
Nicotinamide  
Gastrin  
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  
3D Growth Matrix Components
Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 See all matrix products
Activin A  
EGF  
FGF basic  
FGF-10 See all FGF Ligands
Noggin See all TGF-beta modulators
R-Spondin 1 See all R-Spondins
Wnt-10b See all Wnts
A 83-01 See all TGF-beta/ALK inhibitor
SB 202190 See all p38 MAPK inhibitors
Prostaglandin E2  
Nicotinamide  
Testosterone  
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-2 Stromal cells
Vimentin Stromal cells
References:
Karthaus W.R. et al. (2014) Cell. 159:163.

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  
3D Growth Matrix Components
Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 See all matrix products
FGF basic See all FGF Ligands
Noggin See all TGF-beta modulators
Y-27623 See all ROCK inhibitors
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  
3D Growth Matrix Components
Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 See all matrix products
Activin A  
FGF basic See all FGF Ligands
FGF-4  
Noggin See all TGF-beta modulators
CHIR 99021 See all GSK-3 inhibitors
SB 431542 See all TGF-beta/ALK inhibitor
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  
3D Growth Matrix Components
Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 See all matrix products
BMP-4 See all BMPs
FGF basic See all FGF Ligands
A 83-01  
SB 431542 See all TGF-beta/ALK inhibitor
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.