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. 2022 Dec 16;3(4):101908.
doi: 10.1016/j.xpro.2022.101908. Epub 2022 Dec 6.

Generation of expandable cardiovascular progenitor cells from mouse and human fibroblasts via direct chemical reprogramming

Jia Wang  1 , Qiyuan Wang  2 , Nan Cao  3
Affiliations

Generation of expandable cardiovascular progenitor cells from mouse and human fibroblasts via direct chemical reprogramming

Jia Wang et al. STAR Protoc. .

Abstract

Here, we present a protocol to reprogram mouse and human fibroblasts into expandable cardiovascular progenitor cells (CPCs) via a defined small-molecule treatment. We describe steps to prepare fibroblasts and generate the chemically induced CPCs (ciCPCs), followed by expansion and differentiation of the ciCPCs. These cells can self-renew in the long term, faithfully retaining the CPC phenotype and cardiovascular differentiation capacity. This protocol provides an autologous and expandable cardiovascular cell source, which may find uses in cardiovascular disease modelling, drug discovery, and cardiac cell therapy. For complete details on the use and execution of this protocol, please refer to Wang et al. (2022).1.

Keywords: Cell Biology; Cell Differentiation; Cell culture; Cell-based Assays; Stem Cells.

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Conflict of interest statement

Declaration of interests The authors declare no competing interests.

Figures

None
Graphical abstract
Figure 1
Figure 1
Reprogramming of MEFs into CPCs via defined small-molecule treatment (A) Morphology of 6C-induced cell colonies derived from MEFs at day 14. Scale bars, 100 μm. (B) Immunofluorescence analyses of Gata4 and Isl1 in 6C-induced cell colonies at day 14. Scale bars, 100 μm.
Figure 2
Figure 2
Reprogramming of HFFs into CPCs via defined small-molecule treatment (A) Morphology of 6C-induced cell colonies derived from HFFs at day 26. Scale bars, 100 μm. (B) Immunofluorescence analyses of GATA4 and ISL1 in 6C-induced cell colonies at day 26. Scale bars, 100 μm.
Figure 3
Figure 3
ciCPCs are long-term expandable in fully chemically defined conditions (A) Morphology of P20 mouse ciCPCs. P, passage. Scale bars, 100 μm. (B) Morphology of P12 human ciCPCs. Scale bars, 100 μm. (C) Immunofluorescence analyses of Ki67, Gata4, and Isl1 in P20 mouse ciCPCs. Scale bars, 100 μm. (D) Immunofluorescence analyses of MEF2C and GATA4 in P12 human ciCPCs. Scale bars, 100 μm.
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