Directed differentiation and long-term maintenance of epicardial cells derived from human pluripotent stem cells under fully defined conditions

Xiaoping Bao, Xiaojun Lian, Tongcheng Qian, Vijesh J. Bhute, Tianxiao Han, Sean P. Palecek

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Here, we describe how to efficiently direct human pluripotent stem cells (hPSCs) differentiation into self-renewing epicardial cells in a completely defined, xeno-free system by temporal modulation of regulators of canonical Wnt signaling. Appropriate differentiation-stage-specific application of Gsk3 inhibitor, Wnt inhibitor, and Gsk3 inhibitor (GiWiGi) is sufficient to produce cells expressing epicardial markers and exhibiting epicardial phenotypes with a high yield and purity from multiple hPSC lines in 16 d. Characterization of differentiated cells is performed via flow cytometry and immunostaining to assess quantitative expression and localization of epicardial cell-specific proteins. In vitro differentiation into fibroblasts and smooth muscle cells (SMCs) is also described. In addition, culture in the presence of transforming growth factor (TGF)-β inhibitors allows long-term expansion of hPSC-derived epicardial cells (for at least 25 population doublings). Functional human epicardial cells differentiated via this protocol may constitute a potential cell source for heart disease modeling, drug screening, and cell-based therapeutic applications.

Original languageEnglish (US)
Pages (from-to)1890-1900
Number of pages11
JournalNature Protocols
Volume12
Issue number9
DOIs
StatePublished - Sep 1 2017

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

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