Background Development of vascularized biological scaffolds providing mechanical protection for islets is a challenging objective of modern regenerative medicine. The aim of our study was to develop vascularized endocrine pancreas using decellularized placental cotyledon. Methods/ Materials Human placentas were decellularized by perfusion, using 0.5% sodium dodecyl sulfate. Decellularization was assessed by histological analyses, scanning electron microscopy (SEM) and residual DNA quantification. Glucosaminoglycans (GAG) and hydroxyproline were analyzed. Placental cotyledons were recellularized by a intravascular perfusion of human umbilical vein endothelial cells (HUVECs) during the 24h, followed by intraparenchimal injection of INS-1E cells. Recellularization was assessed by hystological and immunohistochemical methods, and endocrine function was confirmed by glucose stimulated insulin secretion tests performed at different time points. Results Histological staining and SEM showed complete decellularization and well preserved ECM structure. This was confirmed by the absence of residual DNA. Immunostainings of recellularized cotyedons showed insulin expressing islet like structures with intense vascularization confirmed by CD31 staining. Adequate insulin secretion in response to high glucose stimulation was also observed, confirming the functional activity of INS-1E cells. Conclusion These data demonstrate that acellular placental cotyledons seeded with endocrine pancreatic tissue and endothelial cells could be used for functional pancreas bioengineering.