In our previous research, EPCs has been isolated from animal marrow cells and cord blood and improved the angiogenesis performance. Here we explored the effect of over-expression of Wnt3a gene in endothelial progenitor cells (EPCs) and demonstrated a new therapeutic angiogenesis strategy using genetically modified EPCs. As we previously reported, EPCs could be cultivated from animal peripheral blood. The adenovirus vector encoding Wnt3a gene was transfected into EPCs. Overexpression of Wnt3a firstly activated the Wnt/beta-catenin signaling pathway led to increased nuclear translocation of beta-catenin and increased secretion of angiogenic cytokines (vascular endothelial growth factor-a). It enhanced the survival, proliferation, and differentiation of EPCs. What's more, combination of transplantation of EPCs with injection of adenovirus vector encoding Wnt3a gene into the hind limb ischemic model of mouse significantly improved blood flow and tissue capillary density compared with EPCs only treatment group and hind limb ischemic model. Over expression of Wnt3a in the EPCs augmented the in vitro and in vivo angiogenic potency of these cell populations. These data provide evidence that the Wnt3 pathway potentially has a key role in the angiogenic properties of EPCs. Furthermore, the genetic modification of EPCs to stimulate this signaling pathway would promote the efficacy of the therapeutic angiogenesis.