How are MCF10A cells expressed in two dimensional culture?
MCF10A cells expressed markers representing luminal, basal, and progenitor phenotypes in two-dimensional (2D) culture. When grown in suspension culture, MCF10A cells showed low mammosphere-forming ability. Cells in mammospheres and 3D culture expressed both luminal and basal markers.
How is CK5 staining in MCF10A cells?
Around 50% of cells displayed strong staining of CK5, whereas around 10% of cells expressed moderate levels of CK17. Phalloidin staining (F-actin) indicated stress fiber formation in MCF10A cells. Surprisingly, MCF10A cells exhibited completely negative staining of P63 and less than 1% of cells were positive for CK14.
Can a MCF10A cell be a normal progenitor?
Our results show that MCF10A cells may not represent phenotypically normal luminal, basal, or progenitor/stem cells, thus questioning the relevance of MCF10A as a normal mammary epithelial model. This study was approved by the Institutional Review Board (IRB) at Cedars-Sinai Medical Center.
Is the MCF10A a normal breast cell line?
The MCF10A human breast epithelial cell line is arguably the most commonly used normal breast cell model. These cells were derived from benign proliferative breast tissue and spontaneously immortalized without defined factors. They are not tumorigenic and do not express estrogen receptor [ 6 ].
How are MCF10A cells cultured in DMEM / Ham?
MCF10A cells (American Type Culture Collection, Manassas, VA) were cultured in DMEM/Ham’s F-12 (GIBCO-Invitrogen, Carlsbad, CA) supplemented with 100 ng/ml cholera toxin, 20 ng/ml epidermal growth factor (EGF), 0.01 mg/ml insulin, 500 ng/ml hydrocortisone, and 5% chelex-treated horse serum.
Can you grow MCF10A cells without EGF?
(1) I grow MCF10A cells, but have never tried growing them without cholera toxin. (2) I have tried growing them without insulin added and this makes very little dofference (suprisingly). (3) Growing the cells without hydrocortisone did not work. Growing them without EGF did not work.
Where does the MCF 10A cell line come from?
MCF 10A was derived from adherent cells in the population. Soule HD, et al. Isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10. Cancer Res. 50: 6075-6086, 1990. PubMed: 1975513