Abstract
Mammary epithelial cells are widely used as models in mastitis research and as tools for mammalian bioreactors; however, the short lifespan of these cells limits their utility. Several mammal epithelial cell line models have been established; however, the secretion capacity and the bacterial sensitivity of these lines have not been effectively evaluated. In this study, a stable immortalized goat mammary epithelial cell (GMEC) line was constructed by transfection with the SV40 gene. The monoclonal cells were then passaged through more than 50 generations after puromycin selection. The GMEC line was evaluated by reverse transcriptase polymerase chain reaction, the cell cycle, karyotype analysis, detection of apoptosis, Western blotting, and β-casein (CSN2) inducible assays. The GMEC line had a strong proliferation capacity relative to the primary GMECs. GMECs had the same karyotype as the primary cells. The GMEC lines maintained basic biological properties and had estrogen, prolactin, and progesterone receptors as same the primary cells. Additionally, the cells and the cell line could synthesize and secrete β-casein proteins. Finally, the rate of apoptosis of the transfected cells suggested that the cell line could provide a useful tool for signal research and mammary gland bioreactors.
Highlights
Mastitis is characterized by inflammation of the mammary glands induced by mechanical damage and microbial infection and can be classified as subclinical or clinical mastitis [1]
Primary goat mammary epithelial cell (GMEC) were isolated from mammary gland tissue and seeded into Petri dishes
Cell markers were detected by immunofluorescence assays and the red signal exhibited by CK18, ER, progesterone receptor (PR), and prolactin receptor (PRLR) for antibody-positive cells (Figure 1C)
Summary
Mastitis is characterized by inflammation of the mammary glands induced by mechanical damage and microbial infection and can be classified as subclinical or clinical mastitis [1]. Subclinical mastitis in ewes is often induced by bacterial infections [4]. There is a pressing need to find ways to reduce the incidence of mastitis through the production of antimicrobial peptides. Previous study reported production of antimicrobial peptides in mammary tissue is an effective way to reduce incidence of mastitis [7]. This methods usually employed transgenic methods, which generally involve developing a cell model for antibacterial and secretion assays. A stable cell model is necessary for gene editing of animal reproductive traits and the use of the mammary gland as a bioreactor
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