Culturing Breast Cancer and Endothelial Cells

Triple negative breast cancer cell lines, BT20 (CLS Cell Line Service, Eppelheim, Germany) and SUM149PT (Asterand, Royston, UK), and the derivatives SUM149-scr-1, SUM149-sh5-21, and SUM149-sh5-17 cells, were cultured in Dulbecco´s Modified Eagle Medium (DMEM; Gibco Life Technologies, Grand Island, NY, USA) supplemented with 10% FCS. Human umbilical vein cells (HUVEC) were grown in Clonetics Endothelial Growth Medium (EGM; Promocell, Heidelberg, Germany). The medium was changed every 2 days. When cells reached confluency, they were split in a 1:3 ratio. The collection of primary human umbilical vein endothelial cells (HUVEC) was approved by the ethical review board of the Medical Faculty Carl Gustav Carus of the TU Dresden (EK124082003). Primary cultures of HUVEC were isolated using 0.5% collagenase II solution (Worthington Biochemical Corp. Lakewood, NJ, USA) [30 (link)].