
Organoid and 3D Cell Culture Reagents
Organoid and three-dimensional (3D) cell culture have emerged as pivotal systems for understanding human organ development, modeling disease, screening for drug efficacy or toxicity, and investigating personalized medicine. The reagents and protocols needed for differentiation and culturing these advanced multi-cellular in vitro tissues vary by starting cell type, organ and species, and include various formulations of extracellular matrices, organoid culture media and supplements, proteins, small molecules, and antibodies.
This page serves as a reagent and technical resource to help cell biology researchers optimize culture conditions to build robust organoids with high viability. Access protocols for reproducible 3D organoid culture systems and discover new products relevant to your work in organoid research.
3D Organoid Models
Protocols to generate 3D brain organoids from ESCs and iPSCs were first published in 2009. These studies showed that pluripotent stem cells could differentiate into cerebral organoids containing specific cortical regions, neural progenitor populations, and cortical layer patterning. Cerebral organoids have since been employed to uncover evolutionary differences in brain development between species, mechanisms of brain region interconnectivity, and the developmental physiology of normal and diseased brain regions. iPSC-derived organoids show great potential for use in drug discovery as well as modeling neurodegenerative disease and viral brain infection.
Product Name | Catalog # |
2-mercaptoethanol | Various |
Advanced DMEM/F-12 Cell Culture Medium (Thermo Fisher) | 12634010 |
Cultrex Organoid Harvesting Solution | 3700-100-01 |
Cultrex UltiMatrix Reduced Growth Factor Basement Membrane Extract or Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 |
BME001-05 or 3533-010-02 |
GlutaMAX™ (Thermo Fisher) | 35050061 |
Insulin | Various |
N-2 MAX Supplement | AR009 |
N21-MAX Supplement | AR008 |
N21-MAX Vitamin A Free Supplement | AR012 |
Penicillin/Streptomycin | Various |
Recombinant Human FGF basic or Recombinant Human FGF basic Heat Stable |
BT-FGFB or BT-FGFBHS |
Recombinant Human Noggin | 6057-NG |
Y-27632 dihydrochloride (Rho Kinase Inhibitor) | 1254 |
beta-III Tubulin (clone TuJ-1) | Neuronal marker |
EMX1 | Dorsal cortex marker |
EOMES | Intermediate progenitor marker |
FOXG1 | Forebrain marker |
Frizzled-9 | Hippocampus marker |
MAP2 | Neuronal marker |
Neuropilin-2 | Hippocampus marker |
Pax6 | Neural induction marker/ventral cortex marker |
Prox1 | Hippocampus marker |
SOX1 | Neural inductions |
Vimentin | Neuronal marker |
Cancer organoids should be cultured in similar conditions 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 organoid growth medium.
The stomach contains Lgr5+ adult stem cells that can be isolated, cultured, and differentiated in vitro into gastric organoids. Early organoid models elucidated molecular mechanisms underlying gastric development, including signaling pathways that influence fundic or antral gastric epithelium formation. Gastric organoid cultures are powerful models to study normal and diseased gastric physiology as well as more complex life science models for drug discovery and disease modeling.
Human Gastric Organoid Culture Protocol
Product Name | Catalog # |
A 83-01 (ALK5 Inhibitor) | 2939 |
Advanced DMEM/F-12 Cell Culture Medium (Thermo Fisher) | 12634010 |
CHIR 99021 (GSK-3 Inhibitor) | 4423 |
Cultrex UltiMatrix Reduced Growth Factor Basement Membrane Extract or Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 |
BME001-05 or 3533-010-02 |
Cultrex Organoid Harvesting Solution | 3700-100-01 |
Gastrin I (Human) | 3006 |
GlutaMAX (Thermo Fisher) | 35050061 |
HEPES | 3173 |
Human Insulin, Solution | Various |
Human Transferrin | Various |
N-Acetylcysteine | 7874 |
N-2 MAX Supplement | AR009 |
N21-MAX Supplement | AR008 |
Nicotinamide | 4106 |
Penicillin/Streptomycin | Various |
Recombinant Human EGF | 236-EG |
Recombinant Human FGF-10 | 345-FG |
Recombinant Human Noggin | 6057-NG |
Recombinant Human R-Spondin 1 or Recombinant Human Wnt/R-Spondin Agonist |
4645-RS or BT-WRSP1-3 |
Recombinant Human Wnt-3a | 5036-WN |
SB 202190 (p38 MAPK Inhibitor) | 1264 |
Y-27632 dihydrochloride (Rho Kinase Inhibitor) | 1254 |
E-Cadherin | Gastric epithelium |
Lgr5/GPR49 | Gastric stem cell marker |
MUC5AC | Gastric pit mucous cells |
MUC6 | Gland mucous cells |
Pepsinogen C | Chief cells |
Somatostatin | Enteroendocrine cells |
TFF1 | Gastric cells |
TFF2 | Gastric cells |
In vitro generation of cardiac tissue is enabling advancements in drug discovery and toxicity testing, as well as facilitating the engineering of cardiac tissue for regenerative therapies. Various methods have been employed to generate 3D cardiac tissue, including iPSC-derived cardiomyocyte spheroids and bioprinting of cardiac organoids from iPSCs with subsequent differentiation into cardiomyocytes. However, protocol and reagent advancements are still needed to enhance the maturity and complexity of the cardiac tissue.
alpha-Actin | Cardiomyocyte Intracellular Markers |
Cardiac Troponin | Cardiomyocyte Intracellular Markers |
Connexin 43 | Gap Junction Protein Marker |
Flk-1 | Mesodermal Progenitor Surface Marker |
GATA4 | Cardiac Progenitor Intracellular Markers |
HCN4 | Sinoatrial Pacemaker Cell Markers |
MLC2a | Atrial Maturity |
MLC2v | Ventricular Maturity |
MYH6 | Atrial Cardiomyocyte Markers |
MYH7 | Ventricular Cardiomyocyte Markers |
Nanog | iPSC Marker |
NKX2.5 | Cardiac Progenitor Intracellular Markers |
OCT3/4 | iPSC Marker |
PDGF R alpha | Mesodermal Progenitor Surface Marker |
RYR2 | Calcium Handling, Mature Cardiomyocyte Marker |
SCN5a | Transporter Marker on Plasma Membrane |
SERCA2a | Mature Cardiomyocyte Marker |
SOX2 | iPSC Marker |
SSEA-4 | iPSC Marker |
TBX5 | Cardiac Progenitor Intracellular Markers |
Pluripotent stem cell-derived inner ear organoids are rapidly advancing our understanding of inner ear development and physiology. Inner ear organoids have been shown to develop sensory epithelium containing the necessary hair cells, supporting cells, and synaptic-like structures that support auditory or gravitational transduction. These models have great potential for translational research, uncovering molecular and cellular mechanisms that support the regeneration of cochlear and vestibular sensory tissue.
Product Name | Catalog # |
A 83-01 (ALK5 Inhibitor) | 2939 |
Advanced DMEM/F-12 Cell Culture Medium (Thermo Fisher) | 12634010 |
CHIR 99021 (GSK-3 Inhibitor) | 4423 |
Cultrex Organoid Harvesting Solution | 3700-100-01 |
Cultrex UltiMatrix Reduced Growth Factor Basement Membrane Extract or Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 |
BME001-05 or 3533-010-02 |
GlutaMAX (Thermo Fisher) | 35050061 |
N-2 MAX Supplement | AR009 |
N21-MAX Supplement | AR008 |
PD 0325901 (MEK Inhibitor) | 4192 |
Penicillin/Streptomycin | Various |
Recombinant Human BMP-4 | 314-BP |
Recombinant Human FGF basic or Recombinant Human FGF basic Heat Stable |
BT-FGFB or BT-FGFBHS |
Recombinant Human Leukemia Inhibitor Factor (LIF) | 7734-LF |
SB 431542 | 1614 |
The small intestine, large intestine and colon consist of a multicellular epithelium with distinct morphological structures, including villi and invaginated crypt structures. Intestinal crypts house Lgr5+ intestinal adult stem cells that are responsible for the continuous renewal of intestinal epithelium. Intestinal crypts were first utilized to create long-term 3D culture models of the intestine, termed human intestinal organoids (hIO) or epithelial organoids. Intestinal organoid cultures are employed to study normal and diseased physiology, including barrier functions, nutrient uptake, and tissue renewal. In addition, human intestinal organoids can be generated from iPSCs. iPSC-derived hIOs have been utilized as advanced models for gastrointestinal developmental biology, drug toxicity, and personalized medicine applications.
Human Intestinal Organoid Culture Protocol Mouse Enteric Organoid Culture Protocol
Product Name | Catalog # |
A 83-01 (ALK5 Inhibitor) | 2939 |
Advanced DMEM/F-12 Cell Culture Medium (Thermo Fisher) | 12634010 |
Cultrex UltiMatrix Reduced Growth Factor Basement Membrane Extract or Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 |
BME001-05 or 3533-005-02 |
Cultrex Organoid Harvesting Solution | 3700-100-01 |
GlutaMAX (Thermo Fisher) | 35050061 |
HEPES | 3173 |
N-Acetylcysteine | 7874 |
N21-MAX Supplement | AR008 |
Nicotinamide | 4106 |
Penicillin/Streptomycin | Various |
Prostaglandin E2 | 2296 |
SB 202190 (p38 MAPK Inhibitor) | 1264 |
Recombinant Human EGF | 236-EG |
Recombinant Human R-Spondin 1 or Recombinant Human Wnt/R-Spondin Agonist |
4645-RS or BT-WRSP1-3 |
Recombinant Human Noggin | 6057-NG |
Recombinant Human Wnt-3a | 5036-WN |
Y-27632 dihydrochloride (Rho Kinase Inhibitor) | 1254 |
Alkaline Phosphatase | Enterocytes |
ASCL2/Mash | Crypt cell marker |
Chromogranin A | Enteroendocrine cells |
E-Cadherin | Intestinal epithelium |
Lgr5/GPR49 | Intestinal stem cell marker |
Lysozyme | Paneth cells |
MUC2 | Goblet cells |
Synaptophysin | Enteroendocrine cells |
Villin | Enterocytes |
ASCL2/Mash | Crypt cell marker |
Chromogranin A | Enteroendocrine cells |
E-Cadherin | Intestinal epithelium |
EpCAM/TROP-1 | Epithelial cell marker |
EphB2 | Colon crypt cell marker |
Ki-67/MKI67 | Cell proliferation marker |
Lgr5/GPR49 | Intestinal stem cell marker |
Using pluripotent stem cells, kidney organoid culturing protocols have shown the ability to recapitulate the organ’s complex tissue cytoarchitecture, including expression of cellular markers for podocytes, proximal tubules, and distal tubules. Success in cultivating kidney organoids has facilitated research interrogating kidney development, physiology, and mechanisms underlying kidney disease (i.e. chronic kidney disease). In addition, kidney organoid research has demonstrated its potential as a translational method for kidney tissue regeneration.
Product Name | Catalog # |
Advanced DMEM/F-12 Cell Culture Medium (Thermo Fisher) | 12634010 |
CHIR 99021 (GSK-3 Inhibitor) | 4423 |
Cultrex Organoid Harvesting Solution | 3700-100-01 |
Cultrex UltiMatrix Reduced Growth Factor Basement Membrane Extract or Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 |
BME001-05 or 3533-010-02 |
GlutaMAX (Thermo Fisher) | 35050061 |
Holo-Transferrin | 2914-HT |
N-Acetylcysteine | 7874 |
N-2 MAX Supplement | AR009 |
N21-MAX Supplement | AR008 |
Penicillin/Streptomycin | Various |
Recombinant Human Activin A | 11348-AC |
Recombinant Human BMP-2 | 355-BM |
Recombinant Human BMP-4 | 314-BP |
Recombinant Human FGF basic or Recombinant Human FGF basic Heat Stable |
BT-FGFB or BT-FGFBHS |
Recombinant Human FGF-9 | 273-F9 |
Retinoic Acid | 695 |
Y-27632 dihydrochloride (Rho Kinase Inhibitor) | 1254 |
Brachyury | Mesoderm |
Cytokeratin 8 | Uteric bud |
GATA-3 | Uteric bud |
GFR alpha-1/GDNFR alpha-1 | Uteric bud |
HOXB7 | Uteric bud |
LHX-1/LIM1 | Mesoderm |
Pax2 | Uteric bud |
Pax8 | Uteric bud |
Ret | Uteric bud |
The liver is the primary organ system for drug metabolism and detoxification. In this role, it is also highly susceptible to damage from pharmaceuticals and other chemical toxicants. Animal models and traditional in vitro assays modeling liver metabolism often fail to recapitulate the in vivo toxicity of drugs in human patients. Liver organoids, derived from primary tissue or induced pluripotent stem cells, have emerged as more complex and predictive models for hepatotoxicity and drug screening.
Human Liver Organoid Culture Protocol
Product Name | Catalog # |
Advanced DMEM/F-12 Cell Culture Medium (Thermo Fisher) | 12634010 |
A 83-01 (ALK5 Inhibitor) | 2939 |
Cultrex Organoid Harvesting Solution | 3700-100-01 |
Cultrex UltiMatrix Reduced Growth Factor Basement Membrane Extract or Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 |
BME001-05 or 3533-010-02 |
DAPT | 2634 |
Dexamethasone | 1126 |
Forskolin | 1099 |
Gastrin I (Human) | 3006 |
GlutaMAX (Thermo Fisher) | 35050061 |
HEPES | 3173 |
N-Acetylcysteine | 7874 |
N-2 MAX Supplement | AR009 |
N21-MAX Supplement | AR008 |
Nicotinamide | 4106 |
Recombinant Human BMP-7 | 354-BP |
Recombinant Human EGF | 236-EG |
Recombinant Human HGF | 294-HG |
Recombinant Human FGF-10 | 345-FG |
Recombinant Human FGF-19 | 969-FG |
Recombinant Human Noggin | 6057-NG |
Recombinant Human R-Spondin 1 or Recombinant Human Wnt/R-Spondin Agonist |
4645-RS or BT-WRSP1-3 |
Recombinant Human Wnt-3a | 5036-WN |
Y-27632 dihydrochloride (Rho Kinase Inhibitor) | 1254 |
CD24 | Hepatocyte progenitors |
CD44 | Hepatocyte progenitors |
CD133 | Liver stem cells |
Cytokeratin 7 | Centrilobular hepatocytes |
Cytokeratin 19 | Hepatocytes |
EpCAM/TROP-1 | Epithelial cells |
Glutamine Synthetase | Hepatocyte progenitors |
HNF-1 alpha | Hepatocytes |
HNF-4 alpha/NR2A1 | Hepatocytes |
Integrin beta 1/CD29 | Hepatocytes |
Lgr5/GPR49 | Liver stem cells |
Serum Albumin | Hepatocytes |
Transthyretin/Prealbumin | Hepatocytes |
3D cell culture models of the pulmonary system are increasingly utilized to study lung regeneration, model disease (i.e. cystic fibrosis), and investigate mechanisms of viral lung infection (i.e. SARS-CoV-2). While lung organoids were first generated using Lgr5+ stem cells isolated from primary tissue, protocols for culturing iPSC-derived lung organoids have increased the flexibility and accessibility of this model system for use in personalized medicine and drug discovery.
Human Lung Organoid Culture Protocol
Product Name | Catalog # |
A 83-01 (ALK5 Inhibitor) | 2939 |
Advanced DMEM/F-12 Cell Culture Medium (Thermo Fisher) | 12634010 |
Cultrex Organoid Harvesting Solution | 3700-100-01 |
Cultrex UltiMatrix Reduced Growth Factor Basement Membrane Extract or Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 |
BME001-05 or 3533-010-02 |
GlutaMAX (Thermo Fisher) | 35050061 |
HEPES | 3173 |
N-Acetylcysteine | 7874 |
N21-MAX Supplement | AR008 |
Nicotinamide | 4106 |
Penicillin/Streptomycin | Various |
SB 202190 (p38 MAPK Inhibitor) | 1264 |
Recombinant Human KGF/FGF-7 | BT-KGF |
Recombinant Human FGF-10 | 345-FG |
Recombinant Human Noggin | 6057-NG |
Recombinant Human R-Spondin 1 | 4645-RS |
Y-27632 dihydrochloride (Rho Kinase Inhibitor) | 1254 |
ACTTUB | Ciliated cells |
alpha-Smooth Muscle Actin | Smooth muscle cells |
FoxJ1 | Ciliated cells |
HOPX | Type 1 airway epithelial cells |
ID2 | Lung progenitor marker |
Lgr5/GPR49 | Lung stem cells |
NKX2.1 | Lung lineage marker |
p63 | Basal airway cells |
SFTPC | Type 2 airway epithelial cells |
Uteroglobin/SCGB1A1 | Club cells |
Protocols to generate mammary organoids from primary epithelial tissues are helping elucidate the cell fate decisions and molecular mechanisms of mammary gland development, including ductal formation and transformation of milk-producing alveoli. Most importantly, these 3D culture techniques have enabled the cultivation of breast organoids, which are being employed for in vitro drug discovery and personalized drug screening for breast cancer.
Pancreatic organoids have become an informative in vitro model to study pancreatic cancer, exocrine disease, and the basic development of pancreatic ductal epithelium for potential use as regenerative or therapeutic treatment of diabetes. While robust protocols for pancreatic organoid generation using mouse primary pancreatic ductal tissues exist, protocols that support the long-term cultivation of pancreatic organoids from human tissues are still emerging.
Product Name | Catalog # |
A 83-01 (ALK5 Inhibitor) | 2939 |
Advanced DMEM/F-12 Cell Culture Medium (Thermo Fisher) | 12634010 |
Cultrex Organoid Harvesting Solution | 3700-100-01 |
Cultrex UltiMatrix Reduced Growth Factor Basement Membrane Extract or Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 |
BME001-05 or 3533-010-02 |
Gastrin I (Human) | 3006 |
GlutaMAX (Thermo Fisher) | 35050061 |
N-Acetylcysteine | 7874 |
N-2 MAX Supplement | AR009 |
N21-MAX Supplement | AR008 |
Nicotinamide | 4106 |
Penicillin/Streptomycin | Various |
Recombinant Human EGF | 236-EG |
Recombinant Human FGF-10 | 345-FG |
Recombinant Human Noggin | 6057-NG |
Recombinant Human R-Spondin 1 or Recombinant Human Wnt/R-Spondin Agonist |
4645-RS or BT-WRSP1-3 |
Recombinant Human Wnt-3a | 5036-WN |
HNF-6 | Pancreatic progenitors |
Insulin C-Peptide | Beta cells |
NeuroD1 | Beta cells |
Neurogenin-3 | Pancreatic progenitors |
NKX6.1 | Endocrine cells |
PDX-1 | Pancreatic progenitors |
SOX9 | Pancreatic progenitors |
Prostate organoids have rapidly evolved our ability to investigate prostate cancer, its underlying mechanisms, and novel therapeutic strategies for treatment. Prostate organoids, derived from dissociated tissues, adult stem cells, or pluripotent stem cells, are characterized as spheroid structures with differentiated, pseudostratified epithelial cell layers and express functional androgen receptors. Patient‐derived organoids, such as xenograft (PDX)‐prostate organoids, provide useful model for drug discovery and toxicology screening of potential therapeutics. Learn more about prostate cancer screening tools.
Product Name | Catalog # |
A 83-01 (ALK5 Inhibitor) | 2939 |
Cultrex Organoid Harvesting Solution | 3700-100-01 |
Cultrex UltiMatrix Reduced Growth Factor Basement Membrane Extract or Cultrex Reduced Growth Factor Basement Membrane Extract, Type 2 |
BME001-05 or 3533-010-02 |
GlutaMAX (Thermo Fisher) | 35050061 |
N-Acetylcysteine | 7874 |
N-2 MAX Supplement | AR009 |
N21-MAX Supplement | AR008 |
Nicotinamide | 4106 |
Penicillin/Streptomycin | Various |
Prostaglandin E2 | 2296 |
Recombinant Human Activin A | 11348-AC |
Recombinant Human EGF | 236-EG |
Recombinant Human FGF basic or Recombinant Human FGF basic Heat Stable |
BT-FGFB or BT-FGFBHS |
Recombinant Human FGF-10 | 345-FG |
Recombinant Human Noggin | 6057-NG |
Recombinant Human R-Spondin 1 | 4645-RS |
Recombinant Human Wnt-10b | 7196-WN |
SB 202190 (p38 MAPK Inhibitor) | 1264 |
Testosterone | Various |
Androgen R/NR3C4 | Prostate cells |
Cytokeratin 5 | Basal prostate cell |
Cytokeratin 8 | Luminal prostate cell |
Cytokeratin 18 | Prostate cells |
Integrin alpha 6/CD49f | Epithelial stem cells |
Laminin gamma 1 | Stromal cells |
NKX3.1 | Prostate-specific transcription factor |
p63 | Basal prostate cell |
TMPRSS2 | Prostate cells |
TROP-2 | Epithelial stem cells |
Vimentin | Stromal cells |