Effect Of Different Generations, Glucose And Amino Acid On Casein Gene Expression Of The Bovine Mammary Epithelial Cells

In the experiment, lactating Holstein cow’s mammary epithelial cells isolated from the tissue were taken as research subject, and testing effect of different generations of bovine mammary epithelial cells on casein gene expression, on the basis of it, primary (P0) and second generation (P2) were used to effect of glucose (Glc), lysine and methionine influencing on casein gene expression of mammary epithelial cells, respectively. In order to determine the appropriate generation and provide a theoretical basis for related research.1. Using Real-Time PCR detection technology to detect casein gene expression of primary generation (Po) derived from the bovine mammary tissue, the first (P1) to fifth generation (P5); The results showed that expression of three target-genes (αs1-casein, β-casein, κ-casein) was at different levels between the cells of primary generation and passages. The expression in the primary cells was significantly higher than P1-P5cells (P<0.05), whereas P1-P5cells had no significant difference (P>0.05). Moreover, from Po to P5, the expression of αs1-casein, β-casein, κ-casein is gradually decreasing. So, P2orP3BMECs could be used as a trial material.2. Regarding primary (Po) and the second generation (P2) cow’s mammary epithelial cells as a culturing model in vitro, detecting effect of glucose, lysine, and methionine added additional in medium on casein gene expression of bovine mammary epithelial cells. Medium non-treated were seen as control group, added additional glucose treatment as group Ⅰ, lysine and methionine as treatment group Ⅱ, the treatment group Ⅲ embraces lysine, methionie and glucose. The experimental period of24h, with3replicates each of them. Testing casein genes-αs1-casein, β-casein, κ-casein-expression after experimental treatment. Experimental results showed casein gene expression of the primary and second generation cells in3treated groups of bovine mammary epithelial cells of are higher than that of control group, αs1-casein, β-casein gene expression of the treatment group Ⅰ was lower than the treatment group Ⅲ in the primary, but the difference was not significant (P>0.05), group Ⅲ and Ⅰ was significantly higher(P<0.05) than the treatment group Ⅱ and control group; κ-casein in the control group, treatment group Ⅱ, treatment group Ⅰ, treatment groupⅢ progressively presented an increasing tendency, but there was no significant difference (P>0.05); As for the second generation cells, β-casein in treatment group I was lower than that in (P>0.05) treatment groupⅢ, however, the difference was not significant (P<0.05), group Ⅲ and Ⅰ was higher than the treatment group Ⅱ and the control group, treatment group Ⅱ was higher than the control group but the difference was not significant (P>0.05).Both α sl-casein, κ-casein in the control group, treatment group Ⅱ, treatment group Ⅰ, treatment groupⅢ were increased, gradually, but the difference was not significant (P>0.05); αs1-casein, β-casein, κ-casein gene expression in the primary were higher than that of the second generation, but the difference was not significant (P>0.05).To sum up, casein gene expression in bovine mammary epithelial cells culture system combination with glucose, lysine and methionine were higher than added separately of them. After treatment of glucose, lysine and methionine there were no significant difference in casein gene expression between the primary and second generation. Therefore, researchers could conduct relative study choosing the second generation cells not primary anymore.