Combined Growth Factors Enhanced Angiogenic Potential of Cord Blood-derived Mononuclear Cells Transplanted to Ischemic Limbs

Stem cell therapy has recently been limited by poor engraftment and the marginal influence of the administered cells. This study aimed to enhance the survival and angiogenic capacity of human umbilical cord blood (UCB)-derived mononuclear cells (MNCs) and to demonstrate their therapeutic effects on experimental ischemia. A specific culture medium containing five growth factors (Flt-3L, EGF, TPO, FGF and IGF-1) augmented cell proliferation, adhesion potential as well as stimulated MNCs to become progenitor-like cells. In addition, qRT-PCR demonstrated that MNCs cultured with these five growth factors (5f-MNCs) markedly up-regulated multiple angiogenic, arteriogenic and anti-apoptotic factors compared with uncultured MNCs. In an ischemic hindlimb model, the injection of 5f-MNCs prevented limb loss and augmented blood perfusion, capillary density, vascular maturation and angiogenic cytokines in the affected tissues. In addition, the 5f-MNCs exhibited an increased engraftment rate and an endothelial phenotype and stimulated angiogenic factors in ischemic hindlimbs as demonstrated by flow cytometric, immunohistochemical and qRT-PCR analyses. Taken together, these data suggest that 5f-MNCs could be used as a novel therapy for the treatment of ischemic cardiovascular disease due to their increased level of engraftment and angiogenic potential.