Isolation of simian virus 40-transformed human mammary epithelial stem cell lines that can differentiate to myoepithelial-like cells in culture and in vivo.
Simian Virus 40 (SV40) transformation of primary cultures of human mammary epithelial cells has yielded a cloned epithelial-like cell line and a representative, single-cell subclone. Although apparently homogeneous, both cloned cell lines can also yield small numbers of three other cell types. The more-elongated cell type can be obtained directly by replating cells from the medium of the epithelial-like cell cultures or by picking and culturing single cells to form representative lines. Immunofluorescent and immunocytochemical analysis of these cell lines growing on plastic or as tumor-nodules in nude mice for epithelial membrane antigens, various cytokeratins, various actins, laminin, Type IV collagen, the common acute lymphoblastic leukemia antigen (CALLA), and a 135-kDa glycoprotein confirm the epithelial nature of the epithelial-like cells and suggest a myoepithelial origin for the more-elongated cell type. Ultrastructural analysis largely confirms the results, although the myofilamental bundles can be scanty in the growing myoepithelial-like cells. The other two cell types are possibly related to the keratinizing and casein-secreting cells seen in the epithelial tumor-nodules before and after mating the mice, respectively. The myoepithelial-like cells produce 5- to 17-fold more laminin, Type IV collagen, CALLA, and the 135-kDa glycoprotein than the epithelial cells, and all of these antigens are preferentially found on myoepithelial cells in vivo. It is suggested that the SV40-transformed epithelial cell is an immortalized form of human mammary stem cell which can differentiate in culture and in vivo to myoepithelial-like cells.
Rudland
P S
PS
Biochemistry Department, University of Liverpool, England.
Ollerhead
G
G
Barraclough
R
R
D000199:Actins / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D000951:Antigens, Neoplasm; D000954:Antigens, Surface / Q000032:analysis; D001940:Breast / Q000166:cytology*; D002454:Cell Differentiation; D002461:Cell Line, Transformed; D002471:Cell Transformation, Neoplastic; D002472:Cell Transformation, Viral; D003094:Collagen / Q000032:analysis; D004847:Epithelial Cells; D004848:Epithelium / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D007633:Keratins / Q000032:analysis; D007797:Laminin / Q000032:analysis; D008562:Membrane Glycoproteins / Q000032:analysis; D051379:Mice; D008819:Mice, Nude; D018396:Mucin-1; D009368:Neoplasm Transplantation; D009374:Neoplasms, Experimental / Q000473:pathology; D015260:Neprilysin; D013539:Simian virus 40; D013234:Stem Cells / Q000166:cytology*
D000199:Actins; D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D000951:Antigens, Neoplasm; D000954:Antigens, Surface; D007797:Laminin; D008562:Membrane Glycoproteins; D018396:Mucin-1; D007633:Keratins; D003094:Collagen; D015260:Neprilysin