Establishment and characterization of a lactating bovine mammary epithelial cell model for the study of milk synthesis

Authors

  • Ke Zhao,

    1. Institute of Dairy Science, MOE Key Laboratory of Molecular Animal Nutrition, Zhejiang University, No. 268 Kaixuan Road, Hangzhou 310029, Peoples Republic of China
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  • Hong-Yun Liu,

    Corresponding author
    1. Institute of Dairy Science, MOE Key Laboratory of Molecular Animal Nutrition, Zhejiang University, No. 268 Kaixuan Road, Hangzhou 310029, Peoples Republic of China
      email hyliu@zju.edu.cn or liujx@zju.edu.cn
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  • Miao-Miao Zhou,

    1. Institute of Dairy Science, MOE Key Laboratory of Molecular Animal Nutrition, Zhejiang University, No. 268 Kaixuan Road, Hangzhou 310029, Peoples Republic of China
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  • Jian-Xin Liu

    Corresponding author
    1. Institute of Dairy Science, MOE Key Laboratory of Molecular Animal Nutrition, Zhejiang University, No. 268 Kaixuan Road, Hangzhou 310029, Peoples Republic of China
      email hyliu@zju.edu.cn or liujx@zju.edu.cn
    Search for more papers by this author

email hyliu@zju.edu.cn or liujx@zju.edu.cn

Abstract

This study sought to establish anin vitro lactating BMEC (bovine mammary epithelial cell) model, which may maintain the native function for a period of time. Mammary tissues of midlactation Holstein dairy cows were dispersed and cultured in a medium containing insulin, prolactin, hydrocortisone, transferrin, epidermal growth factor and fetal calf serum. After the cells migrating from the tissue reached ∼80% of confluency, the tissues were removed, and secretory epithelial cells were enriched by digesting with 0.25% trypsin repeatedly to remove fibroblasts. The BMEC cells plated on plastic dishes displayed a monolayer, cobblestone, epithelial-like morphology and formed alveoli-like structures and island monolayer aggregates which are the typical characteristics of the mammary epithelial cells. The isolated cells were identified as of epithelial origin by staining with antibody against cytokeratin 18. A one-half logarithmically growth curve and abundant microvilli and cytoplasmic lipid droplets were observed in these cells. The transcription of the αs1 casein gene and synthesis of αs caseins were also detected in the model. Thus, our lactating BMEC model can be an effective model in vitro for studies of milk synthesis in the bovine mammary gland.

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