Cell Lines : An Invasive Human Breast Cancer Essay

931 Words May 12th, 2016 4 Pages
Three cell lines, 4T1, MCF10A and MDA-MB-231, were purchased from ATCC (Manassas, VA) and kept frozen in liquid nitrogen until use. MCF10A (CRL-10317, ATCC) is a non-tumorigenic human breast tissue cell line of epithelial origin. MDA-MB-231 (HTB-26, ATCC) is an invasive human breast cancer triple negative cell line, displaying epithelial-mesenchymal transformation, derived from pleural effusion of a metastatic breast tumor and 4T1 (CRL-2539, ATCC) is a transformed mouse breast cancer cell line derived from a secondary tumor site (www.atcc.org). All the cell cultures, treatment and fixation procedures were carried out in a vertical chamber flow hood to maintain a sterile environment.
Media for cell lines Cell lines were grown at 370C in a humidified 5% CO2 atmosphere (Napco CO2 Incubator, Model #5100, Thermo Scientific). MCF10A cells were grown in DMEM (D2902-WXIL, Lot #SLBM 1834V, Sigma Aldrich, St. Louis) supplemented with 5% horse serum (H-1270, Lot #059K8445, Sigma Aldrich, St. Louis), 20ng/ml EGF (AF-100-15, Peprotech, Rocky Hill, NJ) 0.5 ug/ml hydrocortisone (H0396, Lot #SLBC908N, Sigma Aldrich, St. Louis), 0.1 ug/ml cholera toxin (C-8052, Sigma Aldrich, St. Louis) and 10ug/ml insulin (I-5500, Lot #92K1209, Sigma Aldrich, St. Louis) (Debnath et al., 2003). 4T1 and MDA-MB-231 cell lines were grown in DMEM (D2902-10X1L, Lot #SLBM1834V, Sigma Aldrich, St. Louis) supplemented with 10% FCS (Fetal Calf Serum, SAFC121334, Lot #6J0670) (Huguet et al., 1994).The protocol for…

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