Molecular Nutrition

Culture and Identification of the Bovine Mammary Epithelial Cell Line

  • ZHAN Kang ,
  • GONG Xiaoxiao ,
  • ZUO Xiaoxin ,
  • CHEN Yinyin ,
  • ZHAN Jinshun ,
  • ZHAO Guoqi
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  • College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China

Received date: 2015-03-02

  Online published: 2015-08-13

Abstract

To establish a functional model of bovine mammary epithelial cell line for the study of lactation regulation, pathogenesis of mastitis, this research applied tissue culture to isolate and culture bovine mammary epithelial cells; the limiting dilution method to purify bovine mammary epithelial cells; methyl thiazolyl tetrazolium (MTT) method was used to identify whether the growth curve of bovine mammary epithelial cells was ‘S’ type or not; cytokeratin 18 immunofluorescence was observed to prove that the cells were epithelial type; karyotype analysis was carried out on bovine mammary epithelial cells in passages 10 and 20. The results showed as follows: 1) bovine mammary epithelial cells could be successfully cultured and passaged 20 generations by tissue culture method. 2) At 5 to 6 days of cultivation, the fibroblast cells proliferated rapidly and a small amount of epithelial cells divided. At 8 days of cultivation, bovine mammary epithelial cells proliferated rapidly and formed island-like colonies, and grew like ‘cobblestone’ and ‘slabstone’ as monolayer. 3) Bovine mammary epithelial cells showed positive reaction against cytokeratine 18 antigen. 4) At passages 10 and 20 of cultivation, the cell chromosome number was 60 and the cells had normal diploid karyotype. In conclusion, tissue culture cells are able to obtain stable, functional bovine mammary epithelial cells, but are not immortalized cells.

Cite this article

ZHAN Kang , GONG Xiaoxiao , ZUO Xiaoxin , CHEN Yinyin , ZHAN Jinshun , ZHAO Guoqi . Culture and Identification of the Bovine Mammary Epithelial Cell Line[J]. Chinese Journal of Animal Nutrition, 2015 , 27(8) : 2544 -2550 . DOI: 10.3969/j.issn.1006-267x.2015.08.028

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