Arising technologies of regenerative medicine have made overwhelming the restraints of organs bioengineered in the laboratory possible. In order to simulate the biochemical, spatial and vascular relationships of the native extracellular matrix (ECM), a scaffold has been needed for bioengineered Pancreas. ECM scaffold can be created from all animal organs. Decellularization has been explained lately to generate native ECM scaffolds from compound organs such as the lung, the liver, the heart and more recently the pancreas. In the current study, generating the whole organ through three – dimensional pancreas scaffold using acellular bovine pancreas, has been explained. Previous studies have supported that the protocol used in the current study adequately eliminates cellular material while saving ECM proteins. In this research heparin sulfate was used for the impregnation of the scaffolds. Heparan sulfate proteoglycans (HSPGs) in the ECM bound to several signaling molecules and regulated ligand-receptor interaction, playing an essential role in embryonic development. It was shown that HSPG could arrange the stem cells in asinus and coated basal lamina and make a native morphological shape of acina cells in pancreas scaffold. In the current investigation, it was indicated that HSPG was intensively expressed in pancreatic islet β –cells after 1 week of age in a bovine pancreas.