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Developing a vascular network for tissue engineering constructs is still a challenge. Cell-sheet technology has already brought promising potential in vascularization. However, endothelial cell source for vascularization is limited due to the difficulties in obtaining endothelial cells from peripheral blood or bone marrow. In this study, we differentiated rabbit bone marrow mesenchymal stem cells (rBMSCs) into endothelial like cells (ECs) by adding vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b-FGF) into the culture medium of BMSCs. Then, the induced ECs were seeded on rBMSCs sheets to form vascularized ECs/BMSCs cell sheets. Results show that ECs on the undifferentiated BMSCs sheets formed networks in vitro and in vivo. In vivo results demonstrated that the quantity of vessels in the non-prevascularized constructs was lower than that of prevascularized constructs. Furthermore, the prevascularized rBMSCs sheet constructs induced functional anastomosis with the host vasculature. These results suggested that using BMSCs as an endothelial cell source and cell sheet technology to create prevascularized cell sheet brings a promising potential for tissue regeneration.