title: Functional vascular smooth muscle cells derived from human induced pluripotent stem cells via mesenchymal stem cell intermediates.
abstract: OBJECTIVE
  Smooth muscle cells (SMC) play an important role in vascular homeostasis and disease. Although adult mesenchymal stem cells (MSC) have been used as a source of contractile SMC, they suffer from limited proliferation potential and culture senescence, particularly when originating from older donors. By comparison, human induced pluripotent stem cells (hiPSC) can provide an unlimited source of functional SMC for autologous cell-based therapies and for creating models of vascular disease. Our goal was to develop an efficient strategy to derive functional, contractile SMC from hiPSC.
  
  
  RESULTS
  We developed a robust, stage-wise, feeder-free strategy for hiPSC differentiation into functional SMC through an intermediate stage of multipotent MSC, which could be coaxed to differentiate into fat, bone, cartilage, and muscle. At this stage, the cells were highly proliferative and displayed higher clonogenic potential and reduced senescence when compared with parental hair follicle mesenchymal stem cells. In addition, when exposed to differentiation medium, the myogenic proteins such as α-smooth muscle actin, calponin, and myosin heavy chain were significantly upregulated and displayed robust fibrillar organization, suggesting the development of a contractile phenotype. Indeed, tissue constructs prepared from these cells exhibited high levels of contractility in response to receptor- and non-receptor-mediated agonists.
  
  
  CONCLUSIONS
  We developed an efficient stage-wise strategy that enabled hiPSC differentiation into contractile SMC through an intermediate population of clonogenic and multipotent MSC. The high yield of MSC and SMC derivation suggests that our strategy may facilitate an acquisition of the large numbers of cells required for regenerative medicine or for studying vascular disease pathophysiology.
authors.affiliation: Bioengineering Laboratory, Department of Chemical and Biological Engineering,  University of Buffalo, The State University of New York, 908 Furnas Hall, Amherst, NY 14260-4200,  USA.
authors.forename: Vivek K
authors.initials: VK
authors.lastname: Bajpai
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Mistriotis
authors.forename: Yuin-Han
authors.initials: YH
authors.lastname: Loh
authors.forename: George Q
authors.initials: GQ
authors.lastname: Daley
authors.forename: Stelios T
authors.initials: ST
authors.lastname: Andreadis
mesh_terms: D000199:Actins / Q000378:metabolism; D050156:Adipogenesis; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D049109:Cell Proliferation; D002478:Cells, Cultured; D016922:Cellular Senescence; D020219:Chondrogenesis; D018920:Coculture Techniques; D018507:Gene Expression Regulation, Developmental; D005838:Genotype; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D008840:Microfilament Proteins / Q000378:metabolism; D024510:Muscle Development / Q000235:genetics; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D018995:Myosin Heavy Chains / Q000378:metabolism; D010012:Osteogenesis; D010641:Phenotype; D015854:Up-Regulation; D014661:Vasoconstriction / Q000187:drug effects; D014662:Vasoconstrictor Agents / Q000494:pharmacology; D000096985:Calponins
chemical_list: D000199:Actins; D015415:Biomarkers; D002135:Calcium-Binding Proteins; D008840:Microfilament Proteins; D018995:Myosin Heavy Chains; D014662:Vasoconstrictor Agents; D000096985:Calponins; C541116:ACTA2 protein, human