Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures.
The three-dimensional culture of MCF-10A mammary epithelial cells on a reconstituted basement membrane results in formation of polarized, growth-arrested acini-like spheroids that recapitulate several aspects of glandular architecture in vivo. Oncogenes introduced into MCF-10A cells disrupt this morphogenetic process, and elicit distinct morphological phenotypes. Recent studies analyzing the mechanistic basis for phenotypic heterogeneity observed among different oncogenes (e.g., ErbB2, cyclin D1) have illustrated the utility of this three-dimensional culture system in modeling the biological activities of cancer genes, particularly with regard to their ability to disrupt epithelial architecture during the early aspects of carcinoma formation. Here we provide a collection of protocols to culture MCF-10A cells, to establish stable pools expressing a gene of interest via retroviral infection, as well as to grow and analyze MCF-10A cells in three-dimensional basement membrane culture.
Debnath
Jayanta
J
Department of Cell Biology, Harvard Medical School, 240 Longwood Avenue, Boston, MA 02115, USA. Jayantha_debnath@hms.harward.edu
Muthuswamy
Senthil K
SK
Brugge
Joan S
JS
D001485:Basement Membrane / Q000166:cytology; D001943:Breast Neoplasms; D018929:Cell Culture Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D003094:Collagen; D004338:Drug Combinations; D004847:Epithelial Cells / Q000166:cytology* / Q000502:physiology; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D021621:Imaging, Three-Dimensional; D007797:Laminin; D042361:Mammary Glands, Human / Q000166:cytology*; D011509:Proteoglycans
D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen