title: Routine culturing of normal, dysplastic and malignant human mammary epithelial cells from small tissue samples.
abstract: 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 produce 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.
authors.affiliation: Department of Anatomy, University of British Columbia, Vancouver, Canada.
authors.forename: J T
authors.initials: JT
authors.lastname: Emerman
authors.forename: D A
authors.initials: DA
authors.lastname: Wilkinson
mesh_terms: D000328:Adult; D001769:Blood; D001940:Breast / Q000473:pathology*; D001943:Breast Neoplasms / Q000473:pathology*; D002118:Calcium / Q000008:administration & dosage / Q000494:pharmacology; D002455:Cell Division; D002478:Cells, Cultured; D003593:Cytoplasm / Q000473:pathology; D004848:Epithelium / Q000473:pathology; D005260:Female; D005348:Fibrocystic Breast Disease / Q000473:pathology*; D006801:Humans; D008854:Microscopy, Electron; D008875:Middle Aged; D014407:Tumor Cells, Cultured
chemical_list: D002118:Calcium