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Human mesenchymal stem cells express a myofibroblastic phenotype in vitro: comparison to human cardiac myofibroblasts

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Abstract

Cardiac fibrosis accompanies a variety of myocardial disorders, and is induced by myofibroblasts. These cells may be composed of a heterogeneous population of parent cells, including interstitial fibroblasts and circulating progenitor cells. Direct comparison of human bone marrow-derived mesenchymal stem cells (BM-MSCs) and cardiac myofibroblasts (CMyfbs) has not been previously reported. We hypothesized that BM-MSCs readily adopt a myofibroblastic phenotype in culture. Human primary BM-MSCs and human CMyfbs were isolated from patients undergoing open heart surgery and expanded under standard culture conditions. We assessed and compared their phenotypic and functional characteristics by examining their gene expression profile, their ability to contract collagen gels and synthesize collagen type I. In addition, we examined the role of non-muscle myosin II (NMMII) in modulating MSC myogenic function using NMMII siRNA knockdown and blebbistatin, a specific small molecule inhibitor of NMMII. We report that, while human BM-MSCs retain pluripotency, they adopt a myofibroblastic phenotype in culture and stain positive for the myofibroblast markers α-SMA, vimentin, NMMIIB, ED-A fibronectin, and collagen type 1 at each passage. In addition, they contract collagen gels in response to TGF-β1 and synthesize collagen similar to human CMyfbs. Moreover, inhibition of NMMII activity with blebbistatin completely attenuates gel contractility without affecting cell viability. Thus, human BM-MSCs share and exhibit similar physiological and functional characteristics as human CMyfbs in vitro, and their propensity to adopt a myofibroblast phenotype in culture may contribute to cardiac fibrosis.

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Acknowledgments

We would like to thank Ryan H. Cunnington, Aresh Sepehri, and Steve Wayne for their technical assistance and loans of equipment. This work was supported by funding from the Canadian Institutes for Health Research (New Emerging Team Grant to GT, RCA and DHF; Operating Grant to IMCD), the Heart and Stroke Foundation (grant-in-aid to IMCD) and the St. Boniface Hospital Foundation.

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Authors and Affiliations

  1. Department of Physiology, Faculty of Medicine, Institute of Cardiovascular Sciences, St. Boniface General Hospital Research Centre, University of Manitoba, Winnipeg, MB, Canada

    Melanie A. Ngo, Alison Müller, Yun Li, Shannon Neumann, Ian M. C. Dixon, Rakesh C. Arora & Darren H. Freed

  2. Department of Surgery, Section of Cardiac Surgery, St. Boniface General Hospital Cardiac Sciences Program, University of Manitoba, Winnipeg, MB, Canada

    Rakesh C. Arora & Darren H. Freed

  3. Cardiac Studies, National Research Council - Institute for Biodiagnostics, Winnipeg, MB, Canada

    Ganghong Tian, Rakesh C. Arora & Darren H. Freed

  4. Cardiac Sciences Program, St. Boniface General Hospital, CR3030 Asper Clinical Research Building, 369 Tache Ave, Winnipeg, MB, R2H 2A6, Canada

    Darren H. Freed

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  1. Melanie A. Ngo
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  2. Alison Müller
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  3. Yun Li
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  4. Shannon Neumann
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Correspondence to Darren H. Freed.

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Ngo, M.A., Müller, A., Li, Y. et al. Human mesenchymal stem cells express a myofibroblastic phenotype in vitro: comparison to human cardiac myofibroblasts. Mol Cell Biochem 392, 187–204 (2014). https://doi.org/10.1007/s11010-014-2030-6

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