Functional Enterospheres Derived In Vitro from Human Pluripotent Stem Cells

• Published in: Stem cell reports 2017-09, Vol.9 (3), p.897-912
• Main Authors: Nadkarni, Rohan R., Abed, Soumeya, Cox, Brian J., Bhatia, Sonam, Lau, Jennifer T., Surette, Michael G., Draper, Jonathan S.
• Format: Article
• Language: English
• Subjects: Animals; Bacterial Infections - immunology; Bacterial Infections - pathology; Cell Cycle; Cell Differentiation; Cell Lineage; Cell Proliferation; Cells, Cultured; Endoderm - cytology; Enterocytes - cytology; enterospheres; Epithelial Cells - cytology; Epithelial Cells - metabolism; Fluorescence; Genes, Reporter; Human Embryonic Stem Cells - cytology; human pluripotent stem cells; Humans; Immunity, Innate; innate immunity; intestinal organoids; intestine; in vitro differentiation; Mice; Phenotype; Pluripotent Stem Cells - cytology; Spheroids, Cellular - cytology; Wnt signaling
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2017.07.024

Intestinal organoids derived from human pluripotent stem cells (hPSCs) are valuable in vitro research models that enable simplified access to human gastrointestinal tissues. Here, we report the in vitro generation of enterospheres (hEnS) from hPSC-derived gastrointestinal epithelial precursors. hEnS are cystic spheroids with a simple uniform structure composed entirely of intestinal epithelium. hEnS express markers of mature brush border cells and share a transcriptome profile similar to that of more mature intestinal organoids. Modulation of signaling cues enables control of hEnS growth and differentiation, including long-term propagation. We show that hEnS can be exploited for functional studies: hEnS display an innate immune response when treated with enteric pathogens, and transgenic modification of hEnS with a fluorescence cell-cycle reporter enables hEnS-forming stem cell enrichment. Our work establishes hEnS as an accessible and tractable in vitro modeling system for studying human gastrointestinal biology. •Generated entirely in vitro, hEnS exhibit features of intestinal epithelium•Long-term growth and differentiation of hEnS requires specific media conditions•hEnS elicit an innate immune response after treatment with enteric pathogens•hEnS can be transgenically modified to enrich for stem cells Organoids derived from human pluripotent stem cells (hPSCs) represent an attractive system for investigating human development and disease in vitro. Nadkarni et al. show that hPSC-derived cystic intestinal organoids, or enterospheres, have a uniform epithelial structure, are easily accessible to genetic modification, and contain functional intestinal cell types that provide an in vitro modeling system for studying human gastrointestinal biology.