A more thorough investigation of substrate requirement for cell growth in BME-UV mammary epithelial cells showed that the most important supplements were lactalbumin hydrolysate, hydrocortisone, and insulin because their omission reduced cell proliferation. Currently, the Chinese Holstein cow is the main dairy cow breed in China. However, few reports dealing with mammary epithelial cell function of Chinese Holsteins have been published. With the fast development of the Chinese dairy industry, more attention is being placed on the mechanisms or factors that might affect milk synthesis and quality. Thus, a primary objective of this investigation was to isolate and culture CMECs in vitro so that their potential as a model to study MEC function in this breed of cattle could be evaluated. The isolated CMECs were thoroughly characterized via morphology, chromosomal analysis, immunocytochemistry, RT-PCR, and Western-Blotting analysis. Establishing optimal culture conditions to allow for Testosterone isocaproate protein synthesis in mammary epithelial cells is of importance, as it would more closely mimic the in vivo Velpatasvir system. Previous studies described testing b-casein expression in cultured epithelial cells. Thus, such an approach was considered important in order to test the isolated cell system. We evaluated the protein synthesis ability of the isolated cells through mRNA and protein expression of CSN2, ACACA and BTN1A1. The total RNA and protein were isolated from native mammary tissue, isolated epithelial cells, epithelial cells cultured in induction media, and also in resuscitated epithelial cells. Expression of mRNA was determined by RT-PCR. b-casein protein expression was detected by WesternBlotting as in previous reports. The RT-PCR and WesternBlotting analysis results both confirmed the ability of the isolated cells to synthesize mammary-specific proteins. Furthermore, data showed that the induction media was able to enhance the ability of the isolated epithelial cells to synthesize protein in vitro.In previous studies it was found that the concentration of fetal bovine serum in media had a strong effect on the proliferation of bovine mammary epithelial cells.