Characterization of epithelial phenotypes in mortal and immortal human breast cells

Int J Cancer. 1992 Feb 1;50(3):463-73. doi: 10.1002/ijc.2910500323.


We have previously described the mortal human breast epithelial culture MCF-10M, that was derived from fibrocystic breast tissue, was cultivated in medium with low calcium content for over 2 years, and spontaneously gave rise to the immortal MCF-10 cell line. The emergence of immortalized cells, characterized by growth in conventional calcium levels, from mortal cells has proven to be a reproducible event. Here we report the establishment of a second immortal line from MCF-10M, designated MCF-10-2, and establishment of the MCF-12 immortal line after long-term cultivation of MCF-12M mortal cells from reduction mammoplasty tissue. DNA fingerprinting demonstrated the independent, human origin and lineage of the MCF-10-2 and MCF-12 cell lines. Both lines require cortisol and EGF for maximal growth. The expression in these cultures of in vivo breast epithelial phenotypes was analyzed using 2-dimensional gel Western blots and immunoperoxidase staining with antibodies to cytokeratins and polymorphic epithelial mucin. MCF-10M and MCF-12M retain the cytokeratin profile of the luminal cell (7, 8, 18, 19), and also express cytokeratin 14, found predominantly in basal cells. The immortal lines express a similar profile, except that cytokeratin 19, a component of the fully differentiated luminal cell, is not expressed in the more uniform population seen in MCF-10 and MCF-12, but is retained in the morphologically mixed, less-selected population of MCF-10-2. Epitopes on the polymorphic epithelial mucin, recognized by antibodies HMFG 1, HMFG 2 and SM-3, were detected in the mortal cultures and in the immortal lines, indicating the occurrence of both normal and abnormal mucin processing. MCF-10, MCF-10-2 and MCF-12 cells do not form tumors in nude mice, but appear to organize as duct-like structures before regressing in the 5th week post injection.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Blotting, Western
  • Breast / cytology*
  • Breast / immunology
  • Cell Division
  • Cell Line
  • Cell Survival
  • Clone Cells
  • DNA Fingerprinting
  • Electrophoresis, Gel, Two-Dimensional
  • Epithelial Cells
  • Epithelium / immunology
  • Humans
  • Immunoenzyme Techniques
  • In Vitro Techniques
  • Keratins / chemistry
  • Mice
  • Mice, Nude
  • Polymorphism, Restriction Fragment Length
  • Time Factors


  • Antibodies, Monoclonal
  • Keratins