Feasibility Study On The Home-made Medium Was Used To Culture Bovine Mammary Epithelial Cells

In this experiment, the Bovine Mammary Epithelial Cells was used for the model to assay the effects of Home made-Medium on Biological Function in vitro. The study was conduct from the cell growth curve, the level of cell proliferation, mRNA Transcription of casein gene and the casein synthesis. The purpose is to exam if the Home made-Medium have negative effect on the biological function of BMEC and lay a foundation for further experiments.The content of this study and results as follows:The mammary epithelial cells were obtained with collagenase digestion and milk collection methods from healthy Chinese Holstein cattle and purified by trypsin. Pure epithelial cells were performed by passages culture、cell cryopreservation and recovery. The abtained cells were identificated by morphology, growth characteristics and immunohistochemistry.The results shows that:The growth curve conformed to the rule of "S" sigmoid curve for proliferating cells with a lag-phase, exponential phase, and steady phas. The time for cell went into exponential was at the3rd day and steady phas at the7th day. CK-18immunohistochemical identification results showed a strongly positive. It confirmed that the cells were typical epithelial cells.In this experiment, the effects of Home made-Medium on biological function in BMEC were researched. A completely random design was used. The control group is DMEM/F12medium and the treatment group is Home made-Medium. Each treatment has three replicates and each replicate has three parallel processing. The results show that: The cell activity of BMEC which treated with Home made-Medium has significantly lower than DMEM/F12, but during the period of lag-phase, the activity of BMEC as the same as DMEM/F12medium. The proliferation of BMEC which treated with Home made-Medium has significantly higher than DMEM/F12medium (P＜0.0001). The Home made-Medium increased CSN1S1and CSN3genes expression significantly (P=0.034P=0.015) than DMEM/F12medium, and CSN1S1and CSN3genes expression were increased more than3.57-fold and3.27-fold. Compare DMEM/F12medium with Home made-medium the content of a s-casein and P-casein have no significant(P=0.188P=0.548).