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Reprogramming of fibroblasts into expandable cardiovascular progenitor cells via small molecules in xeno-free conditions

Nature Biomedical Engineering volume 6, pages 403–420 (2022)Cite this article

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Abstract

A major hurdle in cardiac cell therapy is the lack of a bona fide autologous stem-cell type that can be expanded long-term and has authentic cardiovascular differentiation potential. Here we report that a proliferative cell population with robust cardiovascular differentiation potential can be generated from mouse or human fibroblasts via a combination of six small molecules. These chemically induced cardiovascular progenitor cells (ciCPCs) self-renew long-term in fully chemically defined and xeno-free conditions, with faithful preservation of the CPC phenotype and of cardiovascular differentiation capacity in vitro and in vivo. Transplantation of ciCPCs into infarcted mouse hearts improved animal survival and cardiac function up to 13 weeks post-infarction. Mechanistically, activated fibroblasts revert to a plastic state permissive to cardiogenic signals, enabling their reprogramming into ciCPCs. Expanded autologous cardiovascular cells may find uses in drug discovery, disease modelling and cardiac cell therapy.

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Fig. 1: Reprogramming of MEFs into ciCPCs by small molecules.
Fig. 2: ciCPCs are long-term expandable in fully chemically defined xeno-free conditions.
Fig. 3: ciCPCs exhibit a transcriptional programme similar to developing CPCs.
Fig. 4: Long-term expanded ciCPCs robustly differentiate into CMs, SMCs and ECs in vitro.
Fig. 5: Expanded ciCPCs convert into CMs, SMCs and ECs in injured mouse hearts and improve cardiac function after MI.
Fig. 6: Mechanisms underlying 6C-induced CPC reprogramming.
Fig. 7: Reprogramming of HFFs into hciCPCs by 6C.
Fig. 8: Expanded hciCPCs exhibit robust cardiovascular potency both in vitro and in vivo.

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Data availability

The main data supporting the results in this study are available within the paper and its Supplementary Information. Source data for the figures are provided with this paper. The RNA-sequencing data are available from the NCBI GEO database, via the accession number GSE159081. Source data are provided with this paper.

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Acknowledgements

We thank Y. Zhou (University of North Carolina) for technical support and all lab members for helpful discussion. N.C. was funded by the National Key R&D Program of China (2018YFA0109100 and 2018YFA050830), the National Natural Science Foundation of China (92057113, 82061148011 and 31771508), and the Guangdong Innovative and Entrepreneurial Research Team Program (2016ZT06S029). J.W. was funded by the National Key R&D Program of China (2018YFA0109600) and the National Natural Science Foundation of China (81700233). Z.Z. was funded by the National Key R&D Program of China (2019YFA0111500).

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  1. These authors contributed equally: Jia Wang, Shanshan Gu.

Authors and Affiliations

  1. Zhongshan School of Medicine and the Seventh Affiliated Hospital, Sun Yat-Sen University, Guangdong, China

    Jia Wang, Shanshan Gu, Fang Liu, Zihao Chen, He Xu, Zhun Liu, Weisheng Cheng, Linwei Wu, Tao Xu, Zhongyan Chen, Ding Chen, Xuena Chen, Fanzhu Zeng, Zhiju Zhao & Nan Cao

  2. Key Laboratory for Stem Cells and Tissue Engineering (Sun Yat-Sen University), Ministry of Education, Guangdong, China

    Jia Wang, Shanshan Gu, Fang Liu, Zihao Chen, He Xu, Zhun Liu, Weisheng Cheng, Linwei Wu, Tao Xu, Zhongyan Chen, Ding Chen, Xuena Chen, Fanzhu Zeng, Zhiju Zhao & Nan Cao

  3. Department of Histoembryology, Genetics and Developmental Biology, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Reproductive Medicine, Shanghai, China

    Mingliang Zhang

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Contributions

J.W., S.G. and N.C. conceived the project, carried out the experiments and wrote the manuscript. Zihao Chen, W.C. and T.X. performed bioinformatics analysis. F.L., H.X., Z.L., L.W., Zhongyan Chen, D.C., X.C., F.Z., Z.Z. and M.Z. performed data analysis, provided materials or assisted with the experiments.

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Correspondence to Nan Cao.

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Nature Biomedical Engineering thanks Ke Cheng, Timothy Kamp and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

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Supplementary Video 1

Beating cluster generated after MEF-derived ciCPCs were cultured in cardiac differentiation conditions for 7 d.

Supplementary Video 2

Beating cluster generated after TTF-derived ciCPCs were cultured in cardiac differentiation conditions for 7 d.

Supplementary Video 3

Beating cluster generated after HFF-derived ciCPCs were cultured in cardiac differentiation conditions for 14 d.

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Wang, J., Gu, S., Liu, F. et al. Reprogramming of fibroblasts into expandable cardiovascular progenitor cells via small molecules in xeno-free conditions. Nat. Biomed. Eng 6, 403–420 (2022). https://doi.org/10.1038/s41551-022-00865-7

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