Abstract: The effects of different cryopreservation methods of Holstein cow mammary gland epithelial cells on its viability were studied. Primary bovine mammary gland epithelial cells were passing five generations and adjusting the cell density to 1×106cells/mL, and than being cryopreserved by 6 different methods. The viability of cells include mortality, adhering dish rate, apoptosis rate was measured after being recovered. The results showed that the death rate of cell being cryopreserved by treatments 2, 3, 5 was significant different from treatment 1（P＜0.01）. The difference of death rate of cell being cryopreserved by treatments 1, 4, 6 was no significant(P＞0.05). Death rate of cryopreserved cell from treatments 2 and 6 was significant different (P＜0.01). Adhering rate of cryopreserved cell from treatment 4 was significant higher than treatments 2, 5 and 6(P＜0.01). Cell growth state of treatments 1 and 4 were better than others. The results indicated that the treatment 4 was the best method in this study.

Key words: cryopreservation; adhering rate; apoptosis rate; mammary epithelial cell