Pluripotent Stem Cell Facility
Human Pluripotent Stem Cell-Derived Organoids

Training in the Generation of HIOs, aHGOs, HCOs and More

The Wells Lab has reported a method for the generation of complex, three dimensional intestinal tissues (human intestinal organoids; HIOs) in vitro through directed differentiation of human pluripotent stem cells.

These tissues consist of epithelial layers surrounded by mesenchyme, and contain all major intestinal epithelial cell types including absorptive enterocytes and the three secretory lineages (goblet cells, enteroendocrine cells, and paneth cells), crypt-like proliferative zones that express intestinal stem cell markers, and exhibit structural/ultrastructural and functional properties of in vivo human intestine.

Furthermore, HIOs can be transplanted into immunodeficient mice, wherein they adopt adult intestinal-like tissue architecture and undergo significant maturation.

Subsequently, the Wells Lab has reported methods to generate human colonic organoids (HCO), human antral gastric organoids (aHGO), human fundic gastric organoids (fHGO), and esophageal organoids, representing human colon, antral and fundic stomach, and esophagus, respectively.

Coupled with our ability to modify gene expression in hPSCs, organoids represent a highly tractable, physiologically relevant in vitro system for studying human development, homeostasis, and disease.

We work closely with Dr. Wells and the Center for Stem Cell and Organoid Medicine (CuSTOM) to support their pioneering organoid studies, and have developed an improved protocol for generation of small intestinal organoids. This protocol relies on the generation of cryopreservable mid-hindgut endoderm (MHE) that can be thawed and subsequently aggregated for robust and reproducible HIO, aHGO and HCO generation.

The Pluripotent Stem Cell Facility provides hands-on training in the generation of HIOs, aHGOs, and HCOs. We also provide access to a validated cryopreserved bank of WA01/H1-derived MHE, and a service to generate a bank of MHE from any hPSC line. These services are only available to Cincinnati Children's and University of Cincinnati researchers.

The aggregation of cryopreserved MHE generates HIOs.

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