Abstract: To build up the method of isolating and culturing the dairy endometrial epithelial cells for further study, mechanical method and enzymatic isolation method were used in this study.0.1% pronase was used to soak uterine horns for 16 to 20 h at 4 ℃, and a surgical knife was used to scrape cells from dairy endoemtrium. The dairy endometrial epithelial cells were cultured by trypsin digestion method. The cultured cells were identified by keratin antibody through immunohistochemistry, and the proliferation of the third generation were measured by MTT assay.The results showed that morphology of the cultured cells were similar to epithelia cells by phase-contrast microscope. In addition, the eighth generation still maintained the similar morphology and growth state comparing to the primary generation cells, and the cytokeratin-positive cells ration was more than 80%. The methods used in this study were suitable for culturing the endometrial epithelial cells with highly quantity, activity and purity, it could be popularized in related labs.

Key words: dairy endometrial epithelial cell; culture in vitro; pronasedigestion; mechanical method; keratin antibody