Self-Renewing Endodermal Progenitor Lines Generated from Human Pluripotent Stem Cells

Xin Cheng, Lei Ying, Lin Lu, Aline M. Galvão, Jason A. Mills, Henry C. Lin, Darrell N. Kotton, Steven S. Shen, M. Cristina Nostro, John Kim Choi, Mitchell J. Weiss, Deborah L. French, Paul Gadue

The use of human pluripotent stem cells for laboratory studies and cell-based therapies is hampered by their tumor-forming potential and limited ability to generate pure populations of differentiated cell types in vitro. To address these issues, we established endodermal progenitor (EP) cell lines from human embryonic and induced pluripotent stem cells. Optimized growth conditions were established that allow near unlimited (>1016) EP cell self-renewal in which they display a morphology and gene expression pattern characteristic of definitive endoderm. Upon manipulation of their culture conditions in vitro or transplantation into mice, clonally derived EP cells differentiate into numerous endodermal lineages, including monohormonal glucose-responsive pancreatic β-cells, hepatocytes, and intestinal epithelia. Importantly, EP cells are nontumorigenic in vivo. Thus, EP cells represent a powerful tool to study endoderm specification and offer a potentially safe source of endodermal-derived tissues for transplantation therapies.

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