Abstract: To establish and perfect cell culture technique of endometrial epithelial cell (EnEpC)in dairy cows, primary cell was isolated and cultured by tissue cultivation that combined with enzymolysis of collagenase. The endometrial epithelial cell was purified from primary culture cells by different time digestion of trypsase, and EnEpC was identified by immunohistochemical detection of cytokeratin expression. The results demonstrated that flagstone-like epithelial cell was obtained by the adoptive method, and cytokeratin expression was positive in the cells. The purified cells could be subcultured about 10 times, and cells from generation 2 to 8 maintained stably biological characteristics of morphology and proliferation state that were found in primary culture cells.

Key words: endometrial epithelial cell; cell culture; Holstein dairy cows