Abstract
BackgroundWe hypothesized that bone marrow derived Sca-1+ stem cells (BM Sca-1+) transduced with multiple therapeutic cytokines with diverse effects will induce faster angiomyogenic differentiation in the infarcted myocardium.Methods and ResultsBM Sca-1+ were purified from transgenic male mice expressing GFP. Plasmids encoding for select quartet of growth factors, i.e., human IGF-1, VEGF, SDF-1α and HGF were prepared and used for genetic modification of Sca-1+ cells (GFSca-1+). Scramble transfected cells (ScSca-1+) were used as a control. RT-PCR and western blotting showed significantly higher expression of the growth factors in GFSca-1+. Besides the quartet of the therapeutic growth factors, PCR based growth factor array showed upregulation of multiple angiogenic and prosurvival factors such as Ang-1, Ang-2, MMP9, Cx43, BMP2, BMP5, FGF2, and NGF in GFSca-1+ (p<0.01 vs ScSca-1+). LDH and TUNEL assays showed enhanced survival of GFSca-1+ under lethal anoxia (p<0.01 vs ScSca-1+). MTS assay showed significant increased cell proliferation in GFSca-1+ (p<0.05 vs ScSca-1+). For in vivo study, female mice were grouped to receive the intramyocardial injection of 15 μl DMEM without cells (group-1) or containing 2.5×105 ScSca-1+ (group-2) or GFSca-1+ (group-3) immediately after coronary artery ligation. As indicated by Sry gene, a higher survival of GFSca-1+ in group-3 on day4 (2.3 fold higher vs group-2) was observed with massive mobilization of stem and progenitor cells (cKit+, Mdr1+, Cxcr4+ cells). Heart tissue sections immunostained for actinin and Cx43 at 4 weeks post engraftment showed extensive myofiber formation and expression of gap junctions. Immunostaining for vWF showed increased blood vessel density in both peri-infarct and infarct regions in group-3. Infarct size was attenuated and the global heart function was improved in group-3 as compared to group-2.ConclusionsAdministration of BM Sca-1+ transduced with multiple genes is a novel approach to treat infarcted heart for its regeneration.
Highlights
Stem cell based cell therapy offers a potentially therapeutic option for ischemic heart disease [1]
The results show that Bone marrow-derived stem cells (BMSCs) differentiate into cardiomyocytes and vascular cells, and secrete multiple growth factors and cytokines which may mediate endogenous regeneration via activation of resident cardiac stem cells and neovascularization, and reduce apoptosis [6]
The heart cell therapy with BMSC to compensate for loss of functional cardiomyocytes during the ischemic episode may be less meaningful without restoration of the regional blood flow in the ischemic myocardium
Summary
Stem cell based cell therapy offers a potentially therapeutic option for ischemic heart disease [1]. Bone marrow-derived stem cells (BMSCs) have been widely studied for use in cardiac repair due to their favorable properties including multipotency, transdifferentiation, immunomodulation and free from the risks of teratoma formation. The results show that BMSCs differentiate into cardiomyocytes and vascular cells, and secrete multiple growth factors and cytokines which may mediate endogenous regeneration via activation of resident cardiac stem cells and neovascularization, and reduce apoptosis [6]. It would be practical to combine cell transplantation with therapeutic gene delivery to the heart to achieve maximum benefits of stem cell therapy. We hypothesized that bone marrow derived Sca-1+ stem cells (BM Sca-1+) transduced with multiple therapeutic cytokines with diverse effects will induce faster angiomyogenic differentiation in the infarcted myocardium
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