Objective Preparation of human acellular amniotic membrane scaffold and its effect on the proliferation of amniotic membrane-derived stem cells. Methods The amniotic membrane of pregnant women after caesarean section at 38-40 weeks was used. The amniotic membrane cell components were removed by physical and enzymatic digestion after washing，and the acellular amniotic membrane scaffold material was obtained，and coated culture dishes of different concentrations were prepared. The amniotic membrane after the caesarean section was stripped and the stem cells were separated by flow cytometry. The amniotic membrane-derived stem cells were cultured on petri dish coated with acellular amniotic membrane scaffold and the cell proliferation activity was analyzed by Cell Counting Kit-8 (CCK-8)，and compared with other biological materials coated petri dishes. Results The proliferation rate of stem cells on a culture dish coated with acellular amniotic membrane scaffold was significantly increased，which was significantly related to the coating concentration. Compared with commonly used scaffold materials，it has a stronger effect on stem cell proliferation，and the difference is significant. Conclusion Human acellular amniotic membrane scaffold material is expected to become a new type of biomaterial in the field of stem cell research and regenerative medicine.