Multipotent amniotic membrane stem cells have the capacity to mature into many different cell types. Aim of research to study isolation and differentiation of amniotic stem cells into insulin secrecting cells. Method: Isolation of amniotic stem cell by protease and percoll 40.8 percent and 58.5 percent . Identification of amniotic membranes stem cells by Ocr-4. Amniotic stem cells differentiated into insulin-secreting cells in DMEM medium (Dulbecco's Modified Eagle's medium) supplemented with 10 percent fetal calf serum, 10mM nicotinamide, 55 uM Beta-mercaptoethanol, 1 mM sodium pyruvate. Harvest cells at day 0, 1, 7 and 14 after the addition of differentiation factors. Results: The concentration of insulin protein and messenger RNA levels of insulin increased at day 7 and day 14 in culture process. Conclusion: amniotic membrane stem cells can differentiate into insulin-secreting cells in-vitro.