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

被引：42

作者：

Bao, Xiaoping ^{[1
]}

Lian, Xiaojun ^{[1
,2
,3
,4
]}

Qian, Tongcheng ^{[1
]}

Bhute, Vijesh J. ^{[1
]}

Han, Tianxiao ^{[1
]}

Palecek, Sean P. ^{[1
]}

机构：

[1] Univ Wisconsin, Dept Chem & Biol Engn, Madison, WI 53706 USA

[2] Penn State Univ, Dept Biomed Engn, University Pk, PA 16802 USA

[3] Penn State Univ, Dept Biol, University Pk, PA 16802 USA

[4] Penn State Univ, Huck Inst Life Sci, University Pk, PA 16802 USA

来源：

NATURE PROTOCOLS | 2017年 / 12卷 / 09期

基金：

美国国家科学基金会;

关键词：

SMOOTH-MUSCLE-CELLS; PROMOTES CARDIAC DIFFERENTIATION; ENDOTHELIAL PROGENITORS; SMALL-MOLECULE; GENERATION; ACTIVATION; DERIVATION; ORIGIN; LINES;

D O I：

10.1038/nprot.2017.080

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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)-beta 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.

引用

页码：1890 / 1900

页数：11

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