The present invention relates to a novel pancreatic beta cell differentiation method using a combination of Pref-1 / EGF / insulin, wherein the efficiency of proliferation and differentiation of pancreatic stem cells is increased by using Pref-1 / EGF / . Therefore, when the stem cell is transferred to the beta cell using the pancreatic beta cell differentiation method of the present invention, it is possible to develop a graft source capable of improving beta cell loss and dysfunction, which is the main cause of the development and progression of diabetes I have.
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