Culturing Human Glioblastoma and Breast Cancer Cells

The human glioblastoma U87 and U87-CXCR4 (a U87 cell line stably transfected with human CXCR4) cell lines were provided by Dr. Sridhar Nimmagadda.^{35 (link)} HMFs were kindly provided by Dr. Gary Luker. Two wild-type human breast cancer cell lines, MDA-MB-231, and BT-474 were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). HMFs and U87, MDA-MB-231, and BT-474 human cancer cells were cultured in 10% fetal bovine serum (FBS, Sigma, St. Louis, MO, USA) supplemented MEM (Mediatech, Manassas, VA, USA), DMEM (Mediatech), RPMI 1640 (Sigma), and ATCC 46-X (ATCC) media, respectively. U87-CXCR4 human cancer cells were maintained in DMEM medium supplemented with 15% FBS, 1 μg/ml puromycin (Sigma-Aldrich), and 300 μg/ml G418 (Mediatech). Cells were maintained at 37 °C in a humidified atmosphere containing 5% CO₂.