Objective:To isolate,culture and identify SD rat pancreas adult stem cells used the common isolation method with stem cells at present;establish a basis for further studies of pancreas cancer stem cells. Methods:We had chosen the normal SD rat pancreas digesting with collagen enzyme P. Use low serum and no glucose culture medium to purify pancreas cells. Western blot,RT-PCR and ICH to identify nestin in pancreas adult stem cells. We also proved that there was ABCG2(ATP-binding cassette superfamily G member 2)in the cells. Results:Pancreas adult stem cells and the cells expressed nestin were isolated successfully and validated by western blot,RT-PCR and ICH. Conclusion:We were successful to isolate nestin positive stem cells from SD rat pancreas used collagen enzyme P digesting and low serum,no glucose culture medium purifying.