Characterization of epithelial membrane antigen expression in human mammary epithelium by ultrastructural immunoperoxidase cytochemistry.

The suitability of three acrylic resins for the immunoelectron microscopical localization of cell surface and cytoskeletal antigens in surgically excised, immersion fixed human breast cancer, using an immunogold system, has been assessed. Good localization of milk fat globule membrane was achieved with LR White, LR Gold and Lowicryl K11M, although the embedding schedule for LR White had to be modified. The best results were achieved with Lowicryl K11M. Only scanty labelling of actin and cytokeratin was seen in LR White embedded tissue, whereas there was clear localization in LR Gold and Lowicryl K11M embedded samples. Tubulin and alpha-actinin was detected at low level in tissues in the low temperature embedding resins, but not in LR White embedded samples. The morphology of the latter was poorer, and there was greater variability in ultrastructure and labelling. Of the two low temperature embedding resins, Lowicryl K11M gave slightly better results. However, the advantages could be outweighed by the problem incurred in achieving the low temperatures, and by poorer handling properties than LR Gold.

show abstract

“…The distribution of labelling was similar to that described by other workers (Petersen & Van Deurs, 1986;Major et al, 1987).…”

supporting

confidence: 86%

Post embedding immunoelectron microscopy of human breast cancer: a comparison of three acrylic resins

Corcoran

Walker

1990

Histochem J

View full text Add to dashboard Cite

show abstract

“…Normal (4,5). The basolateral surface rests on a basement membrane (BM) and a discontinuous layer of myoepithelial cells (2).…”

mentioning

confidence: 99%

Interaction with basement membrane serves to rapidly distinguish growth and differentiation pattern of normal and malignant human breast epithelial cells.

Petersen

Rønnov‐Jessen

Howlett

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

967

912

View full text Add to dashboard Cite

Normal human breast epithelial cells show a Studies in the rodent have shown that the extracellular matrix regulates the growth and differentiation of normal mammary epithelial cells in vivo and in culture (6-13).The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.Thus far, efforts to study early events in human breast tumor development have been hampered by the lack of adequate means to distinguish between normal and transformed cells in culture and have relied on the criteria of senescence and immortalization (14-19)-phenomena that as yet do not have clear counterparts in vivo and take a considerable amount of time to establish.We asked whether normal human breast epithelial cells could respond to BM and recapitulate certain aspects of their normal growth and differentiation program as has been shown using rodent models (8-12). We show that human mammary epithelial cells can indeed express a normal pattern of growth and differentiation when cultured within a reconstituted BM derived from the Englebreth-Holm-Swarm (EHS) tumor. Furthermore, in response to exogenous BM, the cells basally deposit an endogenous BM. In contrast, carcinoma cell lines and biopsies from primary breast tumors were not capable of responding appropriately to BM nor were they able to deposit an intact endogenous BM.Other investigators have also used an EHS matrix to study human cell lines, including metastatic breast tumor cells (20)(21)(22)(23)(24)(25). These studies have focused on morphology of the cells on an EHS matrix. By including primary tissues and addressing growth as well as three-dimensional architecture in normal and malignant cell lines and primary cultures, we have been able to exploit cell-extracellular matrix interaction as an assay not only to distinguish between normal and malignant breast cells but also possibly to "grade" atypia and malignancy. In addition, the system can be used to define differentiation markers and to delineate early changes in transformation assays or preneoplastic lesions. MATERIALS AND METHODSCell Culture. Primary breast epithelia were prepared from 12 reduction mammoplasties and three breast carcinoma biopsies (two primaries and one lymph node metastasis). The primary specimens were selected, disaggregated, and cultured in serum-free CDM3 medium (26) without Hepes or phenol red in the basal medium (Dulbecco's modified Eagle's medium/F12) and with triiodothyronine at 10 nM. We used two normal breast epithelial cell lines and MCF-1OA (28)], another "normal" line [HBL-100 (29)], and six tumorigenic breast cell lines. These included HMT-3909S13 (30, 31), MCF-7 (including subline 9), ZR75, T47-D, and 33), and CAMA-1 (34). These were cultured as described initially except for MCF-10A, which was cul- tTo whom reprint requests should be sent at the * address.9064

show abstract

“…At the ultrastructural level, the SLC never reaches the acinus lumen and only a fraction of the ULLC does so (Smith and Chepko 2001). Because sialomucin is the most prominent marker of luminal epithelial cells and is exclusively expressed on the apical surface of luminal epithelial cells (Petersen and van Deurs 1986), this would imply that some epithelial cells (not facing the lumen) are sialomucin negative. Nevertheless, there is reason to believe that these cells are indeed full members of the luminal epithelial lineage.…”

mentioning

confidence: 99%

Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties

Guðjónsson¹,

Villadsen²,

Nielsen³

et al. 2002

Genes Dev.

339

View full text Add to dashboard Cite

The epithelial compartment of the human breast comprises two distinct lineages: the luminal epithelial and the myoepithelial lineage. We have shown previously that a subset of the luminal epithelial cells could convert to myoepithelial cells in culture signifying the possible existence of a progenitor cell. We therefore set out to identify and isolate the putative precursor in the luminal epithelial compartment. Using cell surface markers and immunomagnetic sorting, we isolated two luminal epithelial cell populations from primary cultures of reduction mammoplasties. The major population coexpresses sialomucin (MUC

show abstract

Characterization of epithelial membrane antigen expression in human mammary epithelium by ultrastructural immunoperoxidase cytochemistry.

Cited by 29 publications

References 32 publications

Post embedding immunoelectron microscopy of human breast cancer: a comparison of three acrylic resins

Post embedding immunoelectron microscopy of human breast cancer: a comparison of three acrylic resins

Interaction with basement membrane serves to rapidly distinguish growth and differentiation pattern of normal and malignant human breast epithelial cells.

Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties

Contact Info

Product

Resources

About