Serum-Free Growth of Human Mammary Epithelial Cells: Rapid Clonal Growth in Defined Medium and Extended Serial Passage with Pituitary Extract

A serum-free medium with bovine pituitary extract as the only undefined supplement has been developed for long-term culture of human mammary epithelial cells. This medium supports serial subculture of normal cells for 10-20 passages (1:10 splits) without conditioning or special substrates, and it su...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1984-09, Vol.81 (17), p.5435-5439
Main Authors: Hammond, Susan L., Ham, Richard G., Stampfer, Martha R.
Format: Article
Language:English
Subjects:
Animals
Breast - cytology
Breast - physiology
Cattle
Cell Division - drug effects
Cell growth
Cells
Cells, Cultured
Cholera
Culture Media
Cultured cells
Epithelial Cells
Epithelium - physiology
Humans
Keratins
Kinetics
Organoids
Pituitary Gland - physiology
Subcultures
Tissue Extracts - pharmacology
Toxins
Ungulates
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Description
Summary:A serum-free medium with bovine pituitary extract as the only undefined supplement has been developed for long-term culture of human mammary epithelial cells. This medium supports serial subculture of normal cells for 10-20 passages (1:10 splits) without conditioning or special substrates, and it supports rapid clonal growth with plating efficiencies up to 35%. It consists of an optimized basal nutrient medium, MCDB 170, supplemented with insulin, hydrocortisone, epidermal growth factor, ethanolamine, phosphoethanolamine, and bovine pituitary extract. Replacement of pituitary extract with prostaglandin E1and ovine prolactin yields a defined medium that supports rapid clonal growth and serial subculture for three or four passages. Cultures initiated in these media from normal reduction mammoplasty tissue remain diploid and maintain normal epithelial morphology, distribution of cell-associated fibronectin, expression of keratin fibrils, and a low level of expression of milk fat globule antigen. Large cell populations can now be generated and stored frozen, permitting multiple experiments over a period of time with cells from a single donor. These media greatly extend the range of experiments that can be performed both conveniently and reproducibly with cultured normal and tumor-derived human mammary epithelial cells.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.81.17.5435