Endothelial colony forming cells (ECFCs) are the only endothelial progenitor cells (EPCs) capable of acquiring a mature endothelial phenotype. ECFCs are mainly mobilized from bone-marrow to promote vascularization and represent a promising tool for cell-based therapy of severe ischemic diseases. VEGF stimulates the proliferation of peripheral blood-derived ECFCs (PB-ECFCs) through oscillations in intracellular Ca2+ concentration ([Ca2+]i). VEGF-induced Ca2+ spikes are driven by the interplay between inositol-1,4,5-trisphosphate (InsP3)-dependent Ca2+ release and store-operated Ca2+ entry (SOCE). The therapeutic potential of umbilical cord blood-derived ECFCs (UCB-ECFCs) has also been shown in recent studies. However, VEGF-induced proliferation of UCB-ECFCs is faster compared to their peripheral counterpart. Unlike PB-ECFCs, UCB-ECFCs express canonical transient receptor potential channel 3 (TRPC3), that mediates diacylglycerol-dependent Ca2+ entry. The present study aimed to investigate whether the higher proliferative potential of UCB-ECFCs was associated to any difference in the molecular underpinnings of their Ca2+ response to VEGF. We found that VEGF induces oscillations in [Ca2+]i that are patterned by the interaction between InsP3-dependent Ca2+ release and SOCE. Unlike PB-ECFCs, VEGF-evoked Ca2+ oscillations do not arise in the absence of extracellular Ca2+ entry and after pharmacological (with Pyr3 and flufenamic acid) and genetic (by employing selective small interference RNA) suppression of TRPC3. VEGF-induced UCB-ECFC proliferation is abrogated upon inhibition of the intracellular Ca2+ spikes. Therefore, the Ca2+ response to VEGF in UCB-ECFCs is shaped by a different Ca2+ machinery as compared to PB-ECFCs and TRPC3 stands out as a promising target in EPC-based treatment of ischemic pathologies.