Experimental Study On Amniotic Membrane As The Substrate For Human Endometrial Cells In Vitro

Objective:Cells in vitro using amniotic membrane as the substrate and investigate the effective carrier and good methods for cultivation of the endometrial cell.Methods:We prepared human amniotic membrane substrate wuth epithelial cells and human amnion extracellular matrix which removed epithelial cells by trypsin/EDTA.We plant the endometrial cells on these two substrates mentioned above in vitro, then observed and compared the differences of the distribution and cell morphology among cells cultured on the substrates by inverted microscope.Results:Amnion extracellular matrix substrates for hours dispersedly with a ball shape and strong reflected light. After cultured for 24～48 hours, the cells aggregated together and proliferated greatly.The morphology of cells cultured on substrates were similar to those cultured on cell culture dishes. However, cells cultured on were flatter and lined up irregularly. They were stretched and connected closely with each orther. The amniotic membrane was shrinked after 3～4 weeks latter. Cell proliferation rate was decreased and Cytoplasm became darker. After inoculated on the human amnion substrates 18～20 hours, compared to those cultured on human amnion extracellular matrix, human endometrial cells adhered to the substrates and the time of cell attachment were 6-8hours latter, while the morphology, distribution and the time period before apoptosis were similar.Conclusion1. It is feasible to use amniotic membrane as a substrate when culturing endometrial cells.2. Cells cultured on human amnion extracellular matrix substrates and human amnion substrates showed the similar cell morphology and distributions when observed by light microscopic. Cells cultured on human amnion extracellular matrix substrates were much easer to adhesion and had a rapid proliferation rate. 3. Human amnion extracellular matrix maybe a kind of new substrate in research on tissue engineering of endometrium.