Human mammary epithelial cells in culture: differentiation and transformation

Stampfer, Martha R.; Bartley, J. C.

doi:10.1007/978-1-4613-1733-3_1

Cited by 87 publications

(90 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In normal breast epithelial cultures, the gene was detectable in 13/17 instances, but not found in either tumour-derived epithelial cultures or in tumour cell lines derived from breast or bone. This concurs with other studies that have shown its expression in cultured normal, but not malignant, mammary epithelial cells (Yaswen et al, 1990;Stampfer and Yaswen, 1993). It has also been suggested that in normal breast, the gene is down-regulated in vitro (Yaswen et al, 1990), which may explain the lack of expression of NB-1 in four normal epithelial cultures.…”

Section: Genetic Analysissupporting

confidence: 91%

“…Using a modification of a method based on differential centrifugation followed by culture in selective media (Emerson and Wilkinson, 1990;Speirs et al, 1996a), this report describes our experiences in establishing such cultures over a 3-year period and discusses their benefits over cell lines. The cultures have been characterized by a number of criteria including immunocytochemistry and flow cytometry with a range of primary antibodies, RT-PCR to detect the NB-] gene, a calmodulin-like protein of unknown function that is expressed in cultured normal, but not malignant, mammary epithelial cells (Yaswen et al, 1990;Stampfer and Yaswen, 1993) and the presence of increased reductive activity of the steroid-converting enzyme 17p-hydroxysteroid dehydrogenase type I (17-HSD), which is significantly up-regulated in breast tumours (Vermeulen et al, 1986;Peltoketo et al, 1996). (Speirs et al, 1996b).…”

mentioning

confidence: 99%

“…After 24 h media was removed and replaced with OM. Cells were (Yaswen et al, 1990;Stampfer and Yaswen, 1993 …”

mentioning

confidence: 99%

See 2 more Smart Citations

Short-term primary culture of epithelial cells derived from human breast tumours

Speirs

Green²,

Walton³

et al. 1998

Br J Cancer

View full text Add to dashboard Cite

Summary 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-cellenriched 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 17p-hydroxysteroid dehydrogenase type 1. 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.

show abstract

Section: Genetic Analysissupporting

confidence: 91%

mentioning

confidence: 99%

See 1 more Smart Citation

Short-term primary culture of epithelial cells derived from human breast tumours

Speirs

Green²,

Walton³

et al. 1998

Br J Cancer

View full text Add to dashboard Cite

show abstract

“…17 18 This particular phenotype has been detected in a subset of normal breast epithelial cells that grows out of mammoplasty cultures, after most of the early passage cells have senesced. 19 20 The high proliferative capacity of these cells, together with this uncommitted differentiation state, has led many observers to regard them as being derived from a small "stem cell" pool present in the basal layer of the normal gland. 21 In contrast, most normal breast epithelial cells present in the intact gland and in early passage mammoplasty cultures express CK8, CK18 (weakly), and CK19, but not vimentin.…”

mentioning

confidence: 99%

p53 mutation in breast cancer. Correlation with cell kinetics and cell of origin

Megha

Ferrari²,

Benvenuto³

et al. 2002

Journal of Clinical Pathology

View full text Add to dashboard Cite

Aim: Several studies have investigated the expression of the cytokeratins (CKs), vimentin, the epithelial growth factor receptor (EGFR), the oestrogen receptor (ER), and the progesterone receptor (PgR), in breast cancer, but no study has directly compared p53 mutations with these phenotypic and differentiation markers in the same case. The present study was designed to provide some of this information. Methods: The expression of the p53 and bcl-2 proteins was evaluated by immunohistochemistry in relation to phenotypic characteristics and cellular kinetic parameters (mitotic index and apoptotic index) in 37 cases of ductal carcinoma in situ (DCIS) and 27 cases of infiltrating ductal carcinoma (IDC) of the breast. In addition, p53 gene mutation was examined by polymerase chain reaction single strand conformation polymorphism analysis (SSCP). Results: Thirteen cases (eight DCIS and five IDC) showed expression of CK8, CK14, CK18, vimentin, and EGFR, consistent with a stem cell phenotype, whereas 44 cases (27 DCIS and 17 IDC) showed expression of CK8 and CK1, weak or negative expression of CK18, but were negative for vimentin and EGFR, consistent with a luminal cell phenotype. DCIS and IDC cases with a stem cell phenotype were ER/PgR negative and intermediately or poorly differentiated. In contrast, the cases with luminal cell phenotype were ER/PgR positive and well or intermediately differentiated. In addition, intermediately or poorly differentiated cases with a stem cell phenotype showed higher proliferative activity (per cent of MIB-l positive cells) than did intermediately or well differentiated cases with a luminal cell phenotype. Both DCIS and IDC cases with a stem cell phenotype were p53 positive and bcl-2 negative by immunohistochemistry. In IDC, p53 expression was associated with a reduction of both mitotic index and apoptotic index compared with DCIS. Most of the tumours showing a more differentiated phenotype (luminal) were p53 negative and bcl-2 positive. In these cases, cell kinetic parameters increased from DCIS to IDC. These data suggest the existence of subsets of DCIS and IDC that, because of their phenotypic characteristics, could be derived from subpopulations of normal breast cells having different control mechanisms of cell proliferation and neoplastic progression. Conclusions: These results are compatible with the hypothesis that the phenotype of the cell of origin constrains both tumour phenotype and the choice of genetic events; however, the occurrence of p53 mutants by chance during neoplastic transformation cannot be excluded.

show abstract

“…Normal HMEC from specimen 184 have been transformed to immortality following exposure to the chemical carcinogen benzo(a)pyrene (BaP) (6,7). Primary cultures were grown in MM and exposed 2-3 times to 2~g!m1 BaP.…”

mentioning

confidence: 99%

An in vitro Human Mammary Epithelial Model System for Studies of Differentiation and Carcinogenesis

Stampfer¹,

Yaswen²,

Parry³

et al. 1991

Neoplastic Transformation in Human Cell Culture

Self Cite

View full text Add to dashboard Cite

Human mammary epithelial cells in culture: differentiation and transformation

Cited by 87 publications

References 32 publications

Short-term primary culture of epithelial cells derived from human breast tumours

Short-term primary culture of epithelial cells derived from human breast tumours

p53 mutation in breast cancer. Correlation with cell kinetics and cell of origin

An in vitro Human Mammary Epithelial Model System for Studies of Differentiation and Carcinogenesis

Contact Info

Product

Resources

About