title: Differentiation of Human Induced-Pluripotent Stem Cells into Smooth-Muscle Cells: Two Novel Protocols.
abstract: Conventional protocols for differentiating human induced-pluripotent stem cells (hiPSCs) into smooth-muscle cells (SMCs) can be inefficient and generally fail to yield cells with a specific SMC phenotype (i.e., contractile or synthetic SMCs). Here, we present two novel hiPSC-SMC differentiation protocols that yield SMCs with predominantly contractile or synthetic phenotypes. Flow cytometry analyses of smooth-muscle actin (SMA) expression indicated that ~45% of the cells obtained with each protocol assumed an SMC phenotype, and that the populations could be purified to ~95% via metabolic selection. Assessments of cellular mRNA and/or protein levels indicated that SMA, myosin heavy chain II, collagen 1, calponin, transgelin, connexin 43, and vimentin expression in the SMCs obtained via the Contractile SMC protocol and in SMCs differentiated via a traditional protocol were similar, while SMCs produced via the Sythetic SMC protocol expressed less calponin, more collagen 1, and more connexin 43. Differences were also observed in functional assessments of the two SMC populations: the two-dimensional surface area of Contractile SMCs declined more extensively (to 12% versus 44% of original size) in response to carbachol treatment, while quantification of cell migration and proliferation were greater in Synthetic SMCs. Collectively, these data demonstrate that our novel differentiation protocols can efficiently generate SMCs from hiPSCs.
authors.affiliation: Division of Cardiology, Department of Medicine, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.
authors.forename: Libang
authors.initials: L
authors.lastname: Yang
authors.affiliation: Stem Cell Institute, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.
authors.forename: Zhaohui
authors.initials: Z
authors.lastname: Geng
authors.affiliation: Division of Cardiology, Department of Medicine, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.
authors.forename: Thomas
authors.initials: T
authors.lastname: Nickel
authors.affiliation: Division of Cardiology, Department of Medicine, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.|Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, Minnesota, United States of America.
authors.forename: Caitlin
authors.initials: C
authors.lastname: Johnson
authors.affiliation: Division of Cardiology, Department of Medicine, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.
authors.forename: Lin
authors.initials: L
authors.lastname: Gao
authors.affiliation: Stem Cell Institute, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.
authors.forename: James
authors.initials: J
authors.lastname: Dutton
authors.affiliation: Division of Cardiology, Department of Medicine, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.
authors.forename: Cody
authors.initials: C
authors.lastname: Hou
authors.affiliation: Division of Cardiology, Department of Medicine, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.|Stem Cell Institute, University of Minnesota Medical School, Minneapolis, Minnesota, United States of America.|Department of Biomedical Engineering, University of Minnesota, Minneapolis, Minnesota, United States of America.|Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, Minnesota, United States of America.|Department of Biomedical Engineering, School of Medicine, School of Engineering, The University of Alabama at Birmingham, Birmingham, Alabama, United States of America.
authors.forename: Jianyi
authors.initials: J
authors.lastname: Zhang
mesh_terms: D002135:Calcium-Binding Proteins / Q000235:genetics; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D024042:Collagen Type I / Q000235:genetics; D018031:Connexin 43 / Q000235:genetics; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D008840:Microfilament Proteins / Q000235:genetics; D009124:Muscle Proteins / Q000235:genetics; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000235:genetics; D014746:Vimentin / Q000235:genetics; D000096985:Calponins
chemical_list: D002135:Calcium-Binding Proteins; D024042:Collagen Type I; D018031:Connexin 43; D008840:Microfilament Proteins; D009124:Muscle Proteins; D018995:Myosin Heavy Chains; D014746:Vimentin; D000096985:Calponins; C051615:transgelin