Isolation and Characterization of Ovarian, Lung, and Breast Cancer Cell Lines

Human ovarian cancer cell lines A2780 and SKOV3, lung cancer cell lines NCI-H1650 and A549, and the breast cancer cell line MDA-MB-231 were purchased from the American Type Culture Collection (ATCC). These cell lines were last tested by short tandem repeat profiling in September, 2012. The 22 fresh tumors used in this study were categorized as stage III serous ovarian adenocarcinomas. Primary ovarian cancer cells were isolated from freshtumor samples using cancer cell isolation kit (Panomics, CA) according to the manufacturer's instruction and previously published paper [53 (link)]. Then CD133⁺ CSLCs and CD133⁻ NCSLCs were isolated from purified cancer cells by FACS (AC133-PE, mouse IgG, Miltenyi) as described previously [14 (link)]. All studies on the enrolled patients were performed using protocols approved by the institutional review board of the Third Military Medical University and written informed consent was obtained from each patient. To establish stable CSLCs with reduced levels of CCL5, CSLCs were transduced with lentivirus carrying CCL5-shRNA (or GFP-shRNA as the control), as described previously [14 (link)].