Abstract: Objective: To explore the differentiation methods of the feeder-free cultured human embryonic stem cell (hESCs) to human endometrium-like cell. Methods：The hESCs were differentiated in vitro using the co-cultured method and the cytokine-induced method. Flow cytometry was applied to detect the differentiation efficiency of hESCs by human endometrium epithelial marker cytokeratin-18(CK-18) and human endometrium mesenchymal marker vimentin(VIM). Results：The hESCs cultured with the two methods represented the elongated and spindle shape on day7. Flow cytometry analysis on day 8 showed that the differentiated cells were all negative for vimentin and partially positive for CK-18. Compared to the cytokine-induced method, the co-cultured method obtained more endometrium epithelial-like cells [(55.63±10.29)% vs. (13.9±0.26)%, P＜0.05]. Conclusions：hESCs were successfully induced to differentiate to endometrium epithelial-like cells by both the co-cultured method and the cytokine-induced method. Compared to the cytokine-induced method, the co-cultured method can obtain endometrium epithelial-like cells more efficiently.

Key words: Human embryonic stem cells, Induced pluripotentstem cells, Cell differentiation;, Endometrium, Epithelialcells;, Cell culture techniques;, Feeder-free system