skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Serum-free growth of human mammary epithelial cells: rapid clonal growth in defined medium and extended serial passage with pituitary extract

Abstract

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 E/sub 1/ and ovine prolactin yields a defined medium that supports rapid clonal growth and serial subculture for three of four passages. Cultures initiated in these media from normal reduction mammoplasty tissue remain diploid and maintain normal epithelia 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. 31 references, 3 figures, 4 tables.

Authors:
; ;
Publication Date:
Research Org.:
Univ. of Colorado, Boulder, CO (United States)
OSTI Identifier:
5955565
DOE Contract Number:  
AC03-76SF00098
Resource Type:
Journal Article
Journal Name:
Proc. Natl. Acad. Sci. U.S.A.; (United States)
Additional Journal Information:
Journal Volume: 81:21
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; EPITHELIUM; CELL CULTURES; AMINES; CLONE CELLS; CULTURE MEDIA; HYDROCORTISONE; INSULIN; MAMMARY GLANDS; MAN; PITUITARY HORMONES; PROSTAGLANDINS; ADRENAL HORMONES; ANIMAL TISSUES; ANIMALS; BODY; CORTICOSTEROIDS; GLANDS; GLUCOCORTICOIDS; HORMONES; HYDROXY COMPOUNDS; KETONES; MAMMALS; ORGANIC COMPOUNDS; ORGANS; PEPTIDE HORMONES; PREGNANES; PRIMATES; STEROID HORMONES; STEROIDS; TISSUES; VERTEBRATES; 550300* - Cytology

Citation Formats

Hammond, S L, Ham, R G, and Stampfer, M R. Serum-free growth of human mammary epithelial cells: rapid clonal growth in defined medium and extended serial passage with pituitary extract. United States: N. p., 1984. Web. doi:10.1073/pnas.81.17.5435.
Hammond, S L, Ham, R G, & Stampfer, M R. Serum-free growth of human mammary epithelial cells: rapid clonal growth in defined medium and extended serial passage with pituitary extract. United States. https://doi.org/10.1073/pnas.81.17.5435
Hammond, S L, Ham, R G, and Stampfer, M R. 1984. "Serum-free growth of human mammary epithelial cells: rapid clonal growth in defined medium and extended serial passage with pituitary extract". United States. https://doi.org/10.1073/pnas.81.17.5435.
@article{osti_5955565,
title = {Serum-free growth of human mammary epithelial cells: rapid clonal growth in defined medium and extended serial passage with pituitary extract},
author = {Hammond, S L and Ham, R G and Stampfer, M R},
abstractNote = {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 E/sub 1/ and ovine prolactin yields a defined medium that supports rapid clonal growth and serial subculture for three of four passages. Cultures initiated in these media from normal reduction mammoplasty tissue remain diploid and maintain normal epithelia 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. 31 references, 3 figures, 4 tables.},
doi = {10.1073/pnas.81.17.5435},
url = {https://www.osti.gov/biblio/5955565}, journal = {Proc. Natl. Acad. Sci. U.S.A.; (United States)},
number = ,
volume = 81:21,
place = {United States},
year = {Sat Sep 01 00:00:00 EDT 1984},
month = {Sat Sep 01 00:00:00 EDT 1984}
}