title: Cardiomyocyte differentiation of bone marrow-derived Oct-4+CXCR4+SSEA-1+ very small embryonic-like stem cells.
abstract: VSELs are a population of rare Oct-4+CXCR4+ CD133+lin-CD45- cells that are deposited during early embryogenesis in developing organs/tissues as a reserve population of pluripotent stem cells for regeneration. We reported recently that they are mobilized into peripheral blood during acute myocardial infarction (AMI) in mice and humans. However, although freshly isolated VSELs in experimental AMI mouse models improve cardiac function, the number of these cells is limited and an ex vivo expansion strategy is needed to employ these cells more efficiently for cardiac regeneration. The aim of this study was to establish an efficient method to expand ex vivo BM-derived very small embryonic-like stem cells (VSELs) into cardiomyocytes. VSELs, highly purified by FACS from the bone marrow of GFP transgenic mice, were expanded over C2C12 cell line myoblasts and exposed to cardiac differentiating media. The changes in gene expression during cardiac differentiation of VSELs were evaluated by RTQ-PCR, immunostaining and gene array analysis. We developed an efficient, two-step, ex vivo expansion/differentiation model of BM-derived VSELs into cardiomyocytes. First, purified GFP+ VSELs are plated over C2C12 murine cell line myoblasts, where they expand and differentiate into characteristic spheres. Subsequently, cells from these spheres are expanded on cardiac differentiating media into cardiomyocytes. This study demonstrates that murine BM-derived VSELs can be efficiently expanded in vitro and differentiated into cardiomyocytes.
authors.affiliation: Third Division of Cardiology, Medical University of Silesia, Katowice, Poland.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Wojakowski
authors.forename: Michal
authors.initials: M
authors.lastname: Tendera
authors.forename: Magda
authors.initials: M
authors.lastname: Kucia
authors.forename: Ewa
authors.initials: E
authors.lastname: Zuba-Surma
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Milewski
authors.forename: David
authors.initials: D
authors.lastname: Wallace-Bradley
authors.forename: Maciej
authors.initials: M
authors.lastname: Kazmierski
authors.forename: Pawel
authors.initials: P
authors.lastname: Buszman
authors.forename: Eugeniusz
authors.initials: E
authors.lastname: Hrycek
authors.forename: Wieslaw
authors.initials: W
authors.lastname: Cybulski
authors.forename: Grzegorz
authors.initials: G
authors.lastname: Kaluza
authors.forename: Piotr
authors.initials: P
authors.lastname: Wieczorek
authors.forename: Janina
authors.initials: J
authors.lastname: Ratajczak
authors.forename: Mariusz Z
authors.initials: MZ
authors.lastname: Ratajczak
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology; D002454:Cell Differentiation; D048429:Cell Size; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D005260:Female; D016256:Lewis X Antigen / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008954:Models, Biological; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000502:physiology*; D050814:Octamer Transcription Factor-3 / Q000378:metabolism*; D019718:Receptors, CXCR4 / Q000378:metabolism*
chemical_list: C514936:CXCR4 protein, mouse; D016256:Lewis X Antigen; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D019718:Receptors, CXCR4