Routine culturing of normal,dysplastic and malignant human mammary epithelial cells from small tissue samples |
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Authors: | Joanne T Emerman Darcy A Wilkinson |
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Institution: | (1) Department of Anatomy, University of British Columbia, 2177 Wesbrook Mall, V6T 1W5 Vancouver, British Columbia, Canada |
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Abstract: | Summary We compared the growth and morphology of normal, dysplastic and malignant human mammary epithelial cells (HMEC) in medium
containing 5% human serum, a serum-free medium (32) and serum-free medium with a low Ca++ concentration. Tissues were dissociated and epithelial organoids or single cells were seeded onto collagen-coated dishes.
The cells grew in serum-containing medium, but growth of fibroblasts was also stimulated. The serum-free medium consistently
selected for and stimulated the growth of epithelial cells. There was little advantage in reducing the Ca++ concentration to further increase cell yield. This serum-free primary culture system allows us to routinely prouce sufficient
numbers of HMEC from small tissue samples for molecular biological investigations. Furthermore, the maintenance of cells in
a defined medium can provide a system for evaluating the direct effects of factors on gene expression.
This work was supported by a grant from the National Cancer Institute of Canada and funds contributed by Mr. B. T. Wharton
in memory of his wife, Nadia. |
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Keywords: | human mammary epithelial cells primary culture serum-free medium |
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