Abstract
We have evaluated the cardiomyogenic potential of clonal populations of human bone marrow mesenchymal stem cells (BM-MSC). Four rapidly proliferating clones of BM-MSC were obtained from the BM of a healthy donor which were then treated with 5-azacytidine and evaluated for the expression of GATA-4, NKx-2.5, FOG-2, TDGF-1, β-MHC, MEF2D and NPPA genes and cTnT, Desmin and β-MHC proteins. Of the four clones (i) Clone-1 had high expression of GATA-4 (1.89 fold (p<0.05), Nkx2.5 (2.29 fold; p<0.05), FOG2 (2.76 fold; p<0.05), TDGF1 (6.97 fold, p<0.005), βMHC (10.22 fold; p<0.005), MEF-2D (1.91 fold; p<0.005) and NPPA (1.65 fold; p<0.005); (ii) clone-2 had up-regulation of Nkx2.5 (1.98 fold; p<0.05) but down-regulation of rest of the genes; (iii) clone-3 had up-regulation of Nkx2.5 (2.11 fold; p<0.05), TDGF1 (1.88 fold; p<0.05), MEF-2D (1.30 fold; p<0.05) and NPPA (1.21 fold; p<0.05), down regulation of GATA-4 and Fog-2 but no change in βMHC gene; and (iv) clone-4 had up-regulation of MEF-2D (1.17 fold; p<0.05) and down regulation of GATA-4, Nkx2.5 but no change in other genes compared to untreated cells of the clones. At the protein level, clone-1 expressed cTnT, Desmin, and βMHC; clone-2 Desmin only while clones-3 and 4 each expressed cTnT, Desmin, and βMHC. Our data shows that BM-MSC are a heterogenous population of stem cells with sub-populations exhibiting a marked difference in the expression of cardiac markers both at gene and protein levels. This highlights that administering selected sub-populations of BM-MSC with a cardiomyogenic potential may be more efficacious than whole population of cells for cardiac regeneration.
Highlights
Bone marrow-derived mesenchymal stem cells (BM-MSC) are being widely explored for cardiac regenerative therapy[1,2,3], based on their ability to differentiate into cardiomyocytes in vitro and in the damaged myocardium in vivo[4,5,6]
The present study reports that human BM-MSC consist of different subpopulations of stem cells having variable degree of cardiomyogenic potency as revealed by expression of GATA-4, Nkx2.5, FOG2, TDGF1 and βMHC cardiomyogenic genes and cardiac troponin T (cTnT), Desmin and βMHC cardiac structural proteins in 5-azacytidine pretreated single cell derived BM-MSC clones
The differentiation of stem cells into functional cardiomyocytes requires a combinatorial action of multiple transcription factors including GATA-4, Nkx2.5, FOG2, TDGF-1 and β-MHC [17, 18]
Summary
Bone marrow-derived mesenchymal stem cells (BM-MSC) are being widely explored for cardiac regenerative therapy[1,2,3], based on their ability to differentiate into cardiomyocytes in vitro and in the damaged myocardium in vivo[4,5,6]. Their secretome is reported to contribute to the therapeutic effects by paracrine mechanisms[7, 8]. Our hypothesis was that there may be cardiac heterogeneity amongst sub-populations of MSC and infusing populations with optimal cardiomyogenic potential would have a bearing in cardiac regeneration
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