title: Human mesenchymal stem cells express a myofibroblastic phenotype in vitro: comparison to human cardiac myofibroblasts.
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.
authors.affiliation: Department of Physiology, Faculty of Medicine, Institute of Cardiovascular Sciences, St. Boniface General Hospital Research Centre, University of Manitoba, Winnipeg, MB, Canada.
authors.forename: Melanie A
authors.initials: MA
authors.lastname: Ngo
authors.forename: Alison
authors.initials: A
authors.lastname: Müller
authors.forename: Yun
authors.initials: Y
authors.lastname: Li
authors.forename: Shannon
authors.initials: S
authors.lastname: Neumann
authors.forename: Ganghong
authors.initials: G
authors.lastname: Tian
authors.forename: Ian M C
authors.initials: IM
authors.lastname: Dixon
authors.forename: Rakesh C
authors.initials: RC
authors.lastname: Arora
authors.forename: Darren H
authors.initials: DH
authors.lastname: Freed
mesh_terms: D001483:Base Sequence; D024042:Collagen Type I / Q000096:biosynthesis; D017931:DNA Primers; D006801:Humans; D066298:In Vitro Techniques; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D009206:Myocardium / Q000166:cytology*; D058628:Myofibroblasts / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D024042:Collagen Type I; D017931:DNA Primers