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[Cancer Research 50, 6087-6094, September 15, 1990]
© 1990 American Association for Cancer Research

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Ultrastructural and Immunocytochemical Characterization of an Immortalized Human Breast Epithelial Cell Line, MCF-101

Larry Tait, Herbert D. Soule and Jose Russo2

Department of Pathology [L. T., J. R.] and Breast Cancer Program [H. D. S.], Michigan Cancer Foundation, Detroit, Michigan 48201

The human breast epithelial cell line MCF-10 was derived from s.c. mastectomy tissue from a 36-year-old, parous, premenopausal woman with fibrocystic disease. It was initiated as a mortal cell line (MCF-10M), which senesces when transferred serially in 1.05 mM calcium. These cells spontaneously gave origin to two immortal sublines, MCF-10A, or attached cells, and MCF-10F, or floating cells, which have proliferated for more than 4 years in Dulbecco's modified essential medium and Ham's F-12 either with the customary calcium concentration of 1.05 mM (DMEM-H) or in medium containing 0.04 mM calcium or low calcium. Studies reported here indicate that MCF-10 is a mammary epithelial cell line. Electron microscopy showed that both MCF-10A and MCF-10F have characteristics of luminal ductal cells, but not of myoepithelial cells. When grown for more than 1200 days in Dulbecco's modified essential medium-Ham's F-12 and low calcium media, respectively, they maintained their epithelial characteristics, although the concentration of calcium exerted a powerful influence on cell morphology. Cells grown in Dulbecco's modified Eagle's medium-Ham's F12 medium are low cuboidal with numerous desmosomes and short microvilli, whereas those grown in low calcium medium have significantly reduced number of desmosomes, are more spherical, and have greater numbers of microvilli which are longer than those of cells grown in DMEM-H. The breast epithelial origin of these cells was confirmed by immunocytochemical detection of epithelial sialomucins and keratins. The monoclonal antibodies MFA-breast and MC5 and the polyclonal antibody epithelial membrane antigen were used to detect the epithelial sialomucins. Keratins were characterized by using KA-4, K-14, AE1/AE3, and K-19 specific antibodies. It was concluded that MCF-10A and MCF-10F cells are breast epithelial cells and that they represent an important tool for studies of the basic processes of growth and carcinogenesis.

1 These investigations were supported by USPHS Grant CA38921 awarded by the National Cancer Institute, Department of Health and Human Services; a grant from Abbott Laboratories; and an Institutional Grant from the United Foundation of Greater Detroit.

2 To whom requests for reprints should be addressed.

Received 11/27/89. Accepted 6/12/90.




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Copyright © 1990 by the American Association for Cancer Research.