Establishment of Immortalized Cow Mammary Epithelial Cells Expressing Both hTERT and SV40 T

Vectors of pc DNA3. 1-h TERT and pc DNA 3. 1-SV40 T were established. After linearization,they were cotransfected to mammary epithelial cells of Holstein cow,in order to research on the role of h TERT and SV40 T in immortalized mammary epithelial cells in vitro. Both PT-PCR and immunohistochemical assays of cells were carried out. Results showed that the expression of h TERT and SV40 T could effectively prolong the culture time in vitro of mammary epithelial cells,and enhance the cell passage number. The obtained cell line could be expressed normally,indicating that the in vitro cultured mammary epithelial cells expressing both h TERT and SV40 T could effectively prolong cell life without affecting the characteristics of mammary cells.