Rpmi 1640 10 040 cv

Human EOC cell line A2780-CP20 (designated as CP20) was a kind gift from Dr. Anil Sood (MD Anderson Cancer Center, Houston, TX); OV90 was from American Type Culture Collection (Manassas, VA). EGFP (enhanced green fluorescent protein) stably expressing CP20 or OV90 cells were established by transfecting the cells with pcDNA3-EGFP vector (Watertown, MA) with FuGENE 6 Transfection Reagent (E2691, Promega, Madison, WI). All cell lines were cultured in RPMI 1640 (10-040-CV, Corning, NY) with 10% Fetal Bovine Serum (FBS, 16000-044, Life technologies, Carlsbad, CA) and 1% Penn-Strep (15140-122, Life technologies). Primary ovarian CAF TAF18 were isolated and identified in this laboratory [39 (link)], grown in DMEM:F12 (10-090, Corning, NY) with 15% FBS and 1% Penn-Strep, and used up to passage 7. HMEC, a kind gift from Dr. Xin Zhang (OUHSC Stephenson Cancer Center, Oklahoma City, OK), and HUVEC (Lonza, Walkersville, MD) were propagated in Endothelial Cell Growth Medium 2 (EGM, CC-3162) and used up to passage 7 [56 (link)]. When different types of cells were cultured together, the mixed cells were grown in a mixed medium prepared by combining equal volumes of the individual medium for each of the three cell types. All cells were maintained in a humidified 95% air and 5% CO2 atmosphere at 37 °C.