Comparison on Culture Methods of Bovine Mammary Epithelial Cells and Induction of Hormones and Cytokines on &beta;-Casein mRNA Expression

LI Wenqing, WANG Jiaqi, NAN Xuemei, BU Dengpan

doi:10.3969/j.issn.1006-267x.2014.09.021

2014 , Vol. 26 >Issue 9: 2607 - 2614

DOI: https://doi.org/10.3969/j.issn.1006-267x.2014.09.021

Ruminants and Herbivorous Animal Nutrition

Comparison on Culture Methods of Bovine Mammary Epithelial Cells and Induction of Hormones and Cytokines on β-Casein mRNA Expression

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1. State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
2. College of Life Science, Henan Agricultural University, Zhengzhou 450002, China;
3. CAAS-ICRAF Joint Laboratory on Agroforestry and Sustainable Animal Husbandry, Beijing 100193, China;
4. Synergetic Innovation Center of Safety and Nutrition, Northeast Agricultural University, Harbin 150030, China

Received date: 2014-04-08

Online published: 2014-09-10

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Abstract

This experiment was conducted to establish an efficient culture method for bovine mammary epithelial cells, and to research the induction effect of hormones and cytokines on β-casein (CNS2) mRNA expression. Bovine mammary epithelial cells were cultured by tissue mass culture method and mechanical disruption method, respectively. Cells were purified according to different sensitivity to trypsin of fibroblasts and bovine mammary epithelial cells. The growth curve of purified cells was detected by cell counting assay method. The expression of keratin 18 was detected by immunofluorescent histochemistry staining method. CNS2 mRNA expression induced by 8 kinds of hormones and cytokines combination mediums was measured by real time quantified PCR (RT-qPCR). The results showed that vigorous bovine mammary epithelial cells could be obtained by mechanical disruption method. Cell migration speed of mechanical disruption method was faster than that of tissue mass culture method. The growth curve showed a typical ‘S’ type. The expression of cytokeratin 18 was found in purified cells and the 20th generation of cells. The supplementation of 100 ng/mL insulin-like growth factor Ⅰ (IGF-Ⅰ) significantly improved CNS2 mRNA expression in mammary epithelial cells (P<0.05). It is concluded that the mechanical disruption method used in the present experiment is an efficient method for bovine mammary epithelial cells, and 100 ng/mL IGF-Ⅰ shows the best induction effect on CNS2 mRNA expression.

Key words： primary cell culture; mammary epithelial cell; hormone; CNS2 mRNA

Cite this article

LI Wenqing, WANG Jiaqi, NAN Xuemei, BU Dengpan . Comparison on Culture Methods of Bovine Mammary Epithelial Cells and Induction of Hormones and Cytokines on β-Casein mRNA Expression[J]. Chinese Journal of Animal Nutrition, 2014 , 26(9) : 2607 -2614 . DOI: 10.3969/j.issn.1006-267x.2014.09.021

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