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.

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

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.

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

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

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.

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.