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Establishment and characterization of a dairy goat mammary epithelial cell line with human telomerase (hT‐MECs)
Authors:Jun Luo  Wei Wang  Abiel B. Haile  Huifen Xu  Jun Li
Affiliation:College of Animal Science and Technology, Northwest A&F University, , Yangling, Shaanxi, China
Abstract:Although research on dairy goat mammary gland have referred extensively to molecular mechanisms, research on lines of dairy goat mammary epithelial cells (MECs) are still rare. This paper sought to establish an immortal MEC line by stable transfection of human telomerase. MECs from a lactating (45 days post‐parturition) Xinong Saanen dairy goat were cultured purely and subsequently transfected with a plasmid carrying the sequence of human telomerase. Immortalized MECs by human telomerase (hT‐MECs) exhibited a typical cobblestone morphology and activity and expression levels of telomerase resembled that of MCF‐7 cells. hT‐MECs on passage 42 grew vigorously and ‘S’ sigmoid curves of growth were observed. Moreover, hT‐MECs maintained a normal chromosome modal number of 2n = 60, keratin 8 and epithelial membrane antigen (EMA) were evidently expressed, and beta‐casein protein was synthesized and secreted. Beta‐casein expression was enhanced by prolactin (P < 0.05). Lipid droplets were found in hT‐MECs, and messenger RNA levels of PPARG, SREBP, FASN, ACC and SCD in hT‐MECs (passage 40) were similar to MECs (passage 7). In conclusion, the obtained hT‐MEC line retained a normal morphology, growth characteristics, cytogenetics and secretory characteristics as primary MECs. Hence, it can be a representative model cell line, for molecular and functional analysis, of dairy goat MECs for an extended period of time.
Keywords:dairy goat  human telomerase  mammary epithelial cell
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