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Isolation, characterization, and EGFP expression in the buffalo (Bubalus bubalis) mammary gland epithelial cell line

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Abstract

This study was aimed to establish a buffalo mammary epithelial cells (BuMECs) line and maintain it for long-term by subculturing. BuMECs isolated from lactating buffalo mammary glands were cultured on a collagen matrix gel. BuMECs expressed significant amounts of the epithelial cell specific marker cytokeratin 18 as determined by immunohistochemistry. The BuMECs displayed monolayer, cobble-stone morphology, and formed lumen-, dome-, and duct-like structures. Furthermore, they were capable of synthesizing CSN2, BLG, ACACA, and BTN1A1, showed viability after thawing and expressed milk protein genes. The enhanced green fluorescent protein gene was transferred successfully into the BuMECs using lipofection method and the transfected cells could be maintained for long-term in culture by subculturing.

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Acknowledgments

This work was supported by grant BT/PR15035/AAQ/01/462/2011 from the Department of Biotechnology, Government of India, New Delhi.

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Authors and Affiliations

  1. Embryo Biotechnology Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal, 132001, India

    Ramakant Kaushik, Karn Pratap Singh, Manoj Kumar Singh, Radhey Shyam Manik, Prabhat Palta, Suresh Kumar Singla & Manmohan Singh Chauhan

  2. Department of Bioscience and Biotechnology, Banasthali University, Rajasthan, 304 022, India

    Archana Kumari

Authors
  1. Ramakant Kaushik
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  2. Karn Pratap Singh
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  3. Archana Kumari
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  4. Manoj Kumar Singh
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  5. Radhey Shyam Manik
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  6. Prabhat Palta
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  7. Suresh Kumar Singla
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Correspondence to Manmohan Singh Chauhan.

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Editor: T. Okamoto

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Kaushik, R., Singh, K.P., Kumari, A. et al. Isolation, characterization, and EGFP expression in the buffalo (Bubalus bubalis) mammary gland epithelial cell line. In Vitro Cell.Dev.Biol.-Animal 49, 1–7 (2013). https://doi.org/10.1007/s11626-012-9557-1

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  • DOI: https://doi.org/10.1007/s11626-012-9557-1

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