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Immortalization of bone marrow‐derived porcine mesenchymal stem cells and their differentiation into cells expressing cardiac phenotypic markers
Author(s) -
Moscoso Isabel,
RodriguezBarbosa JoseIgnacio,
BarallobreBarreiro Javier,
A Patricia,
Domenech Nieves
Publication year - 2012
Publication title -
journal of tissue engineering and regenerative medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.835
H-Index - 72
eISSN - 1932-7005
pISSN - 1932-6254
DOI - 10.1002/term.469
Subject(s) - mesenchymal stem cell , cd90 , cd44 , microbiology and biotechnology , biology , cell culture , cell therapy , immortalised cell line , bone marrow , stem cell , phenotype , cellular differentiation , cell , immunology , cd34 , genetics , gene
Abstract Mesenchymal stem cells (MSCs) may be among the first stem cell types to be utilized in the clinic for cell therapy, because of their ease of isolation and extensive differentiation potential. Using a porcine model, we have established several cell lines from MSCs to facilitate in vitro and in vivo studies of their potential use for cellular therapy. Bone marrow‐derived primary MSCs were immortalized using the pRNS‐1 plasmid. We obtained four stable immortalized cell lines that exhibited higher proliferative capacities than the parental cells. All four cell lines displayed a common phenotype similar to that of primary mesenchymal cells, characterized by constitutively high expressions of CD90, CD29, CD44, SLA I and CD46, while CD172a, CD106 and CD56 were less expressed. Remarkably, treatment with 5‐azacytidine‐stimulated porcine MSCs lines to differentiate into cells that were positive for cardiac phenotypic markers, such as α ‐actin, connexin‐43, sarcomeric actin, serca‐2 and, to a lesser extent, desmin and troponin‐T. These porcine MSC lines will be valuable biological tools for developing strategies for ex vivo expansion and differentiation of MSCs into a specific lineage. Copyright © 2011 John Wiley & Sons, Ltd.