Immortalized Human Mammary Epithelial Cells-GFP (MCF-12A)

Immortalized Human Mammary Epithelial Cells‑GFP (MCF‑12A‑GFP) is a non‑tumorigenic, spontaneously immortalized breast‑epithelial cell line engineered for expression of enhanced green fluorescent protein for live‑cell imaging. Parental MCF‑12A cells were obtained from reduction mammoplasty tissue from a white, nulliparous woman with fibrocystic breast disease that contained focal intraductal hyperplasia. They were established by long‑term culture in serum‑free, low‑calcium medium, a method of immortalization that maintains an adherent, epithelial morphology.

GFP‑tagged derivatives are usually produced by lentiviral or plasmid transduction of a GFP cassette followed by antibiotic selection (puromycin or G418) to select for stable, brightly fluorescent clones. GFP expression allows for real‑time monitoring of cell proliferation, migration and three‑dimensional morphogenesis, and allows for co‑culture studies in which normal mammary epithelium can be distinguished from malignant counterparts. Functionally, MCF‑12A cells are estrogen‑receptor negative, express a combination of epithelial (CK8, CK14, E‑cadherin) and mesenchymal markers, and form lumenized acini or duct‑like structures in collagen or Matrigel matrices. MCF‑12A cells are widely used as a "normal" control in breast‑cancer research, in studies of ETS transcription‑factor‑mediated transformation and in radiobiology studies of low‑energy X‑rays.