Human amniotic epithelial cells culture in vitro and the research of Stem cell characteristics

Objective To establish in vitro culture procedure of human amniotic epithelial cells (hAECS) and to explore the biological characteristics. Methods Human placenta were deliveried from third-trimester healthy mothers. The amnion layer was mechanically peeled off of the chorion, washed several times and splitted in 1 mm × 1 mm. The amnion membrane was incubated with 0.05% trypsin containing 0.53 mM EDTA4Na and then cultured. The hAECS were obtained from the amniotic membrane by the digestion of trypsin. The stem cell markers (0CT-4、Nanog、SSEA-4) were examined by the Immunofluorescence, Western. Result The amniotic epithelial cells express 0CT-4、Nanog、SSEA-4, which is the marker of the stem cells according to immunofluorescence staining and Westernblot results. Conclusion the hAECS express the marker of embryonic stem cell, which means it can be used as a new source of stem cells.