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Growth and differentation of rat mammary epithelial cells cultured in serum-free medium

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Abstract

A new serum-free defined medium was developed that supports the growth of normal rat mammary epithelial cells. Mammary organoids from the glands of female F344 rats were cultured in a serum-free medium. Monolayer culture colonies developed within a week and remained viable for months in culture. Upon subculture of one-week-old primary colonies, almost the same morphology of colonies was developed. The scrape loading/dye transfer technique showed that most of colonies that developed in a serum-free medium containing EGF, human transferrin, insulin, and hydrocortisone (basal serum-free medium, BSFM) failed to show cell-cell communication. However, colonies cultured in BSFM supplemented with prolactin, E2, and progesterone (complete hormone serum-free medium, CHSFM) showed cell-cell communication at 14 days of primary culture or of subculture. By flow cytometry with FITC-PNA and PE-anti-Thy-1.1 monoclonal antibody, we distinguished four RMEC subpopulations in cultures in both media: Thy-1.1+ cells, PNA+ cells, cells negative to both reagents and cells positive to both reagents. It is likely that combined prolactin, cortisol, and insulin in CHSFM stimulate terminal differentiation of clonogenic cells.

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Authors and Affiliations

  1. College of Pharmacy, Pusan National University, 609-735, Pusan, Korea

    Dong Yeum Kim, Byung-Hak Jhun, Kyung Hee Lee, Seung Chul Hong & Nam Deuk Kim

  2. Research Institute of Drug Development, Pusan National University, 609-735, Pusan, Korea

    Dong Yeum Kim, Byung-Hak Jhun, Kyung Hee Lee, Seung Chul Hong & Nam Deuk Kim

  3. Department of Human Oncology, K4/330 CSC, University of Wisconsin Comprehensive Cancer Center, 600 Highland Avenue, 53792, Madison, Wisconsin, U.S.A.

    Kelly H. Clifton

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  1. Dong Yeum Kim
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Kim, D.Y., Jhun, BH., Lee, K.H. et al. Growth and differentation of rat mammary epithelial cells cultured in serum-free medium. Arch. Pharm. Res. 20, 297–305 (1997). https://doi.org/10.1007/BF02976190

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