title: Short-term primary culture of epithelial cells derived from human breast tumours.
abstract: As experimental models for breast cancer, most studies rely on established human breast cancer cell lines. However, many of these lines were established over 20 years ago, many from pleural effusions rather than the primary tumour, so the validity of using them as representative models is questionable. This paper describes our experiences, over a 3-year period, in establishing short-term epithelial-cell-enriched preparations from primary breast tumours based on differential centrifugation followed by culture in selective media. Epithelial cells were successfully cultured from 55% of samples, but culture success did not appear to be correlated with tumour histology, stage, grade or node status. Epithelial cell-enriched cultures were immunopositive for broad-spectrum cytokeratin and epithelial membrane antigen (EMA). Positivity for keratin 19 confirmed that the cultures contained tumour-derived cells, which additionally showed significantly higher activity of the reductive pathway of the steroid-converting enzyme 17beta-hydroxysteroid dehydrogenase type I. That the cultures contained tumour and not normal epithelial cells was further substantiated by the complete absence of the calmodulin-like gene NB-1 in tumour-derived cultures; this is only associated with normal breast epithelia. Eighty-five per cent of cultures established from oestrogen receptor (ER)-positive tumours expressed ER in vitro; this was functional in 66% of cultures, although ER-positive phenotype was gradually lost over time. In conclusion, epithelial cells can be isolated and maintained as short-term cultures from primary breast tumours irrespective of histopathological or clinical details, providing a model system with a greater biological and clinical relevance than breast cancer cell lines.
authors.affiliation: Department of Medicine, University of Hull, UK.
authors.forename: V
authors.initials: V
authors.lastname: Speirs
authors.forename: A R
authors.initials: AR
authors.lastname: Green
authors.forename: D S
authors.initials: DS
authors.lastname: Walton
authors.forename: M J
authors.initials: MJ
authors.lastname: Kerin
authors.forename: J N
authors.initials: JN
authors.lastname: Fox
authors.forename: P J
authors.initials: PJ
authors.lastname: Carleton
authors.forename: S B
authors.initials: SB
authors.lastname: Desai
authors.forename: S L
authors.initials: SL
authors.lastname: Atkin
mesh_terms: D015067:17-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D001943:Breast Neoplasms / Q000737:chemistry / Q000201:enzymology / Q000473:pathology*; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division; D002469:Cell Separation; D004847:Epithelial Cells / Q000737:chemistry / Q000166:cytology* / Q000201:enzymology; D005260:Female; D006801:Humans; D009363:Neoplasm Proteins / Q000378:metabolism; D010641:Phenotype; D011960:Receptors, Estrogen / Q000032:analysis
chemical_list: D009363:Neoplasm Proteins; D011960:Receptors, Estrogen; D015067:17-Hydroxysteroid Dehydrogenases; C044256:3 (or 17)-beta-hydroxysteroid dehydrogenase