Abstract: In this research paper, purified goat mammary epithelial cell line was obtained by culturing dispersed cells routinely and purifying them with time-controlled trypsinization. Results indicated that: large numbers of dispersed mammary cells could be obtained by digesting goat mammary tissue with 150 U/mL collegenaseⅠand 150 U/mL hyaluronidase for 4 h at 37 ℃, which is the feasible method for collection goat mammary cells. Adding hydrocortisone, insulinlike growth factor-I (IGF-I), epithelial growth factor (EGF) and insulin-transferrin-seuim (ITS) to RPMI1640 or DEME/F12 medium, which supplied with 10% fetal bovine serum and antibiotics, was benefical to the goat mammary cell growth significantly. The light microscope structure and perspective electron microscope ultrastructure of goat mammary cell indicated that: goat mammary cells formed island monolayer aggregates and dome-like structure. The seventh generation mammary cell cytoplasms had plenty of secretive vesicles and Golgi apparatus, which showed that the seventh generation mammary had vigorous secretive activity and there wasn’t any sign of apoptosis.