Effects of Self-compounding DMEM/F12 Medium on Biological Function in Primary Cultured Bovine Mammary Epithelial Cells

Abstract

Abstract: The purpose of this study was to exam if the self-compounding medium had negative effects on the biological function of bovine mammary epithelial cells and offered foundation for further experiments.The differences between self-compounding medium and DMEM/F12 medium in this experiment were compared based on determination of cell proliferation, lactating genes expression and casein synthesis. The proliferation of BMEC cultured in self-compounding medium was higher significantly than that of DMEM/F12 medium（P < 0.0001）. The CSN1S1 and CSN3 genes expression of BMEC cultured in self-compounding medium increased significantly（P < 0.05）and up-regulated 3.57-fold and 3.27-fold, respectively. There were no significance of the CSN1S2, CSN2 gene expression and αS-casein, β-casein synthesis（P >0.05）.Compared with DMEM/F12 medium, self-compounding medium had no negative effects on the lactating genes expression and the casein synthesis, and the further study on effects of different amino acid patterns on lactating genes expression and casein synthesis.

Key words: Self-compounding DMEM/F12 medium, Lactating gene expression, Casein synthesis, Cell proliferation

Zhang Xingfu, Du Ruiping, Gao Min, Ao Changjin, Lu Dexun, Cao Qina. Effects of Self-compounding DMEM/F12 Medium on Biological Function in Primary Cultured Bovine Mammary Epithelial Cells[J]. Biotechnology Bulletin, 2013, 0(3): 114-119.

References

[1] Wheeler TT, Callaghan MR, Davis SR, et al. Milk protein synthesis, gene expression, and hormonal responsiveness in primary cultures of mammary cells from lactating sheep[J]. Exp Cell Res, 1995, 217 (2):346-354.
[2] Ehmann VL, Peterson WD. To grow mouse mammary epithelial cell in culture[J]. J Cell Boil, 1984, 98(3):1026-1032.
[3] Deves R, Boyd CA. Transporters for cationic amino acids in animal cells :discovery, structure, and function [J]. Physiol Rev, 1998, 78(2):487-545.
[4] Backwell FR, Bequette BJ, Wilson D, et al. Evidence for the utilization of peptides for milk protein synthesis in the lactating dairy goat in vivo[J]. American Journal of Physiology, 1996, 271(4):955- 960.
[5] Kimball SR, Shantz LM, Horetsky RL, Jefferson LS. Leucine regulates translation of specific mRNAs in L6 myoblasts through mTORmediated changes in availability of eIF4E and phosphorylation of ribosomal protein S6[J]. J Biol Chem, 1999, 274 :11647-11652.
[6] Jobgen WS, Fried SK, Fu WJ, et al. Regulatory role for the argininenitric oxide pathway in metabolism of energy substrates[J]. J Nutr Biochem, 2006, 17 :571-588.
[7] Wu HH, Zhao XJ, Zhang CQ, Liu JX. Establishment of mammary tissue culture :an in vitro mode for bovine lactation[J]. Journal of Animal and Feed Sciences, 2004, 13(Suppl.1):575-578.
[8] Wang X, Campbell LE, Miller CM, Proud CG. Amino acid availability regulates p70 S6 kinase and multiple translation factors[J]. Biochem J, 1988, 334 :261-267.
[9] Wang X, Beugnet A, Murakami M, et al. Distinct signaling events downstream of mTOR cooperate to mediate the effects of amino acids 2013年第3期119 张兴夫等:自制DMEM/F12 培养基对奶牛乳腺上皮细胞生物学功能的影响 and insulin on initiation factor 4E-binding proteins[J]. Mol Cell Biol, 2005, 25 :2558-2572.
[10] Yokota T, Kanamoto R, Hayashi S. c-myc mRNA is stabilized by deprivation of amino acids in primarycultured rat hepatocytes[J]. J Nutr Sci Vitaminol(Tokyo), 1995, 41 :455-463.
[11] Kimball SR, Jefferson LS. Control of protein synthesisby amino acid availability[J]. Curr Opin Clin Nutr Metab Care, 2002, 5 :63- 67.
[12] Ebner KE, Hoover CR, Hageman EC, Larson BL. Cultivation and properties of bovine mammary cell cultures[J]. Experimental Cell Research, 1961, 23 :373-381.
[13] Thordarson G, Ogren L, Day JR, et al. Mammary gland development and alpha-lactalbum in production in hypophysectomized, pregnantmice [J]. Biology Reproduction, 1989, 40(3):517-525.
[14] Wu HH, Zhao XJ, Zhang CQ, Liu JX. Establishment of mammary tissue culture :an in vitro mode for bovine lactation[J]. Journal of Animal and Feed Sciences, 2004, 13(Suppl.1):575-578.
[15] Liu HY, Yang JY, Wu HH, et al. Effects of methionine and its ratio to lysine on expression of αs1-casein gene in cultured bovine mammary epithelial cells[J]. Journal of Animal and Feed Sciences, 2007, 16(Suppl.2):330-334.
[16] Wang C, Liu JX. Effects of ratio of lysine to methionine in metabolizable protein on lactation performance of dairy cows [C]. Proceeding of the 13th Animal Science Congress of the Asian-Australian of Animal Production Societies, 2008.