Immortalized dairy cow mammary epithelial cell line, construction method and applications thereof

Abstract

The invention discloses an immortalized dairy cow mammary epithelial cell line, a construction method, and applications thereof. In the provided construction method, Chinese Holstein cow epithelial cells are taken as the host cells, retrovirus carriers with SV-40-large T antigen are used to infect host cells, and finally a strain of immortalized dairy cow mammary epithelial cell line that can perform stable passage is screened out. The microbial preservation number of the strain is CGMCC No.8707. The constructed immortalized dairy cow mammary epithelial cell line has a stable property, the cell morphology is similar to that of primary cells after 40 generations, and the cells grow faster than the primary cells, can characteristically express mammary epithelial cell specific proteins, and maintain the synthesis functions of milk fat and milk protein. The immortalized dairy cow mammary epithelial cell line can be used as a cell model which is widely used in the fields such as milk component synthesis, milk secretion mechanism, research on laticiferous cell apoptosis and mammary gland degradation mechanism, and the like, and can be used as a mammary gland bioreactor to produce bioactive proteins.

Description

technical field [0001] The invention relates to an immortalized cell line, in particular to an immortalized dairy cow mammary gland epithelial cell line and its construction method and application, and belongs to the field of construction and application of the immortalized dairy cow mammary gland epithelial cell line. Background technique [0002] Cow mammary gland epithelial cells have special functions of synthesizing and secreting milk, and can maintain their characteristics under in vitro culture conditions. The research on cow mammary epithelial cells has been more than half a century. Cow mammary gland epithelial cells can be used to study the expression of milk protein and milk fat genes and the mechanism of milk secretion. As a cell model of mammary gland bioreactor to produce valuable bioactive proteins, it is a research tool with great application value. However, there are great difficulties in the in vitro culture of dairy cow mammary epithelial cells, and the ...

AI Technical Summary

Problems solved by technology

BMGH+HM does not have the polygonal morphology of typical epithelial cells, but has a long and thin shape with long protrusions, but can synthesize cytokeratin and desmosomal proteins; BMGE+H has typical epithelial cell morphology and can synthesize cytokeratin , but cannot form domes or bubble-like structures, and cannot synthesize vimentin

[0005] Although there are many reports on the methods of effectively establishing cell lines at home and abroad, the number and types of mammary epithelial cell lines are far from enough, and many mammary epithelial cell lines are generally unstable during use and have morphological changes. Differentiation or aging after multiple passages, loss of typical characteristics of glandular epithelial cells, etc., need to be improved

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