Cell Culture and Authentication Protocols

MCF-10A cells and their derivatives MCF-ErbB2 and MCF-MekDD cells were provided by M. Reginato (Drexel University, USA) [16 (link)]. MCF10A cells were authenticated as published [11 (link)]. Lack of mycoplasma contamination in all cells was established as published [11 (link)]. BT-474 (American Type Culture Collection (ATCC), Manassas, VA, USA), BT474TR and BT474T cells were cultured as published [11 (link)]. Generation of BT-474TR cells is published [17 (link)]. To generate tumorigenic BT474T cells, BT474 cells were implanted orthotopically into the mammary fat pad of severe combined immunodeficiency mouse, and resulting tumors were serially passaged into new hosts over a three-year period [18 (link)]. AU-565 (ATCC), SKBR3 (ATCC) and 293 T cells (provided by A. Stadnyk, Dalhousie University) were cultured as published [11 (link)]. To detach the cells from the ECM, they were plated in suspension culture above a layer of 1% sea plaque agarose polymerized in respective culture medium not containing any additional ingredients.