title: A Novel Selectable Islet 1 Positive Progenitor Cell Reprogrammed to Expandable and Functional Smooth Muscle Cells.
abstract: Disorders affecting smooth muscle structure/function may require technologies that can generate large scale, differentiated and contractile smooth muscle cells (SMC) suitable for cell therapy. To date no clonal precursor population that provides large numbers of differentiated SMC in culture has been identified in a rodent. Identification of such cells may also enhance insight into progenitor cell fate decisions and the relationship between smooth muscle precursors and disease states that implicate differentiated SMC.  In this study, we used classic clonal expansion techniques to identify novel self-renewing Islet 1 (Isl-1) positive primitive progenitor cells (PPC) within rat bone marrow that exhibited canonical stem cell markers and preferential differentiation towards a smooth muscle-like fate. We subsequently used molecular tagging to select Isl-1 positive clonal populations from expanded and de novo marrow cell populations. We refer to these previously undescribed cells as the PPC given its stem cell marker profile, and robust self-renewal capacity. PPC could be directly converted into induced smooth muscle cells (iSMC) using single transcription factor (Kruppel-like factor 4) knockdown or transactivator (myocardin) overexpression in contrast to three control cells (HEK 293, endothelial cells and mesenchymal stem cells) where such induction was not possible. iSMC exhibited immuno- and cytoskeletal-phenotype, calcium signaling profile and contractile responses similar to bona fide SMC. Passaged iSMC could be expanded to a scale sufficient for large scale tissue replacement.  PPC and reprogramed iSMC so derived may offer future opportunities to investigate molecular, structure/function and cell-based replacement therapy approaches to diverse cardiovascular, respiratory, gastrointestinal, and genitourinary diseases that have as their basis smooth muscle cell functional aberrancy or numerical loss. Stem Cells 2016;34:1354-1368.
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Elizabeth C
authors: EC
authors: Turner
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Chien-Ling
authors: CL
authors: Huang
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Neha
authors: N
authors: Sawhney
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Kalaimathi
authors: K
authors: Govindarajan
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Anthony J P
authors: AJ
authors: Clover
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Kenneth
authors: K
authors: Martin
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Tara C
authors: TC
authors: Browne
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Derek
authors: D
authors: Whelan
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Arun H S
authors: AH
authors: Kumar
authors: Department of Physiology, University College Cork, Biosciences Institute, College Road, Cork, Ireland.
authors: John J
authors: JJ
authors: Mackrill
authors: Molecular Medicine Program, Mayo Clinic and Foundation, 200 First St, Rochester, Minnesota, 55905.
authors: Shaohua
authors: S
authors: Wang
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Jeffrey
authors: J
authors: Schmeckpeper
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Alessia
authors: A
authors: Stocca
authors: Department of Physiology, University College Cork, Biosciences Institute, College Road, Cork, Ireland.
authors: William G
authors: WG
authors: Pierce
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Anne-Laure
authors: AL
authors: Leblond
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Liquan
authors: L
authors: Cai
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Donnchadh M
authors: DM
authors: O'Sullivan
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Chirlei K
authors: CK
authors: Buneker
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Janet
authors: J
authors: Choi
authors: Alimentary Pharmabiotic Centre (APC), Biosciences Institute, University College Cork, Cork, Ireland.
authors: John
authors: J
authors: MacSharry
authors: Molecular Medicine Program, Mayo Clinic and Foundation, 200 First St, Rochester, Minnesota, 55905.
authors: Yasuhiro
authors: Y
authors: Ikeda
authors: Molecular Medicine Program, Mayo Clinic and Foundation, 200 First St, Rochester, Minnesota, 55905.
authors: Stephen J
authors: SJ
authors: Russell
authors: Centre for Research in Vascular Biology (CRVB), Biosciences Institute, University College Cork, Cork, Ireland.
authors: Noel M
authors: NM
authors: Caplice
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D002454:Cell Differentiation; D049109:Cell Proliferation; D000066673:Cell Self Renewal; D002469:Cell Separation; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D002999:Clone Cells; D020868:Gene Silencing; D005822:Genetic Vectors / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000378:metabolism; D060850:LIM-Homeodomain Proteins / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000378:metabolism; D009687:Nuclear Proteins / Q000378:metabolism; D010641:Phenotype; D011916:Rats, Inbred F344; D019098:Telomerase / Q000378:metabolism; D015534:Trans-Activators / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D000099251:Myocardin
chemical_list: D049452:Green Fluorescent Proteins; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D060850:LIM-Homeodomain Proteins; D009687:Nuclear Proteins; D019098:Telomerase; D015534:Trans-Activators; D014157:Transcription Factors; D000099251:Myocardin; C000715467:KLF4 protein, human; C000715469:Klf4 protein, rat; C063843:insulin gene enhancer binding protein Isl-1
keywords: Differentiation; Progenitor cells; Reprogramming; Smooth muscle cells