Organoid and 3D Culture Reagents

Organoids

Organoid and 3D Cell Culture Reagents

What Are Organoids Used For?

Organoid and three-dimensional (3D) cell culture are emerging 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 to culture these advanced multi-cellular in vitro tissues vary by organ and species, as well as whether they are being generated from tissue-specific adult stem cells or induced pluripotent stem cells (iPSCs).

This page serves as a reagent and technical resource to help researchers build robust and consistent organoid cultures designed to provide you with a central location to access protocols, view webinars, stay up to date on organoid recipes and blogs, and discover new products relevant to your work in organoid research. Navigate below to find information for culturing organoids from all tissue types.

Read more about the development and future of organoids for research.

 

 

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How Are Organoids Made?

 

Intestinal Organoids

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.
 

Gasteroids

Stomach epithelial organoids

The stomach contains Lgr5+ adult stem cells that can be isolated, cultured, and differentiated in vitro into gastric organoids. Early organoid models elucidated molecular mechanism 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 models for drug discovery and disease modeling.

 

Lung Organoids

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.

Brain Organoids

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.

 

Liver Organoids

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.

Pancreatic Organoids

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.

 

Kidney Organoids

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.

Heart Organoids

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 with iPSCs that are subsequently differentiated into cardiomyocytes. However, protocol and reagent advancements are still needed to enhance the maturity and complexity of the cardiac tissue.

 

Mammary Organoids

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.

Inner Ear Organoids

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.

 

Prostate Organoids

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 xenograft (PDX)‐prostate organoids provide useful model for drug discovery and toxicology screening of potential therapeutics. Learn more about prostate cancer screening tools.

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.

References:

Fang, Y. et al. (2017) SLAS Discov. 22:456.

Kelm, J.M. et al. (2003) Biotechnol. Bioeng. 83:173.

Fatehullah, A. et al. (2016) Nat. Cell Biol. 18:246.

Shamir, E.R. et al. (2014) Nat. Rev. Mol. Cell Biol. 15:647.

Takasato, M. et al. (2015) Nature 526:564.

Zhu, R. et al. (2014) Stem Cell Res. Ther. 5:117.

Li, Y. et al. (2014) Organogenesis 10:159.

Birgersdotter, A. et al. (2005) Semin. Cancer Biol. 15:405.

Wilson, S.S. et al. (2015) Mucosal Immunol. 8:352.

Lancaster, M. et al. (2013) Nature 501:373.