C 22110

HUVEC were isolated from human umbilical cords as described previously [47 (link)]. Cells were seeded on gelatin coated dishes and cultured in endothelial cell growth medium (Promocell, C-22110) containing supplements (Promocell, C-39210) and 2% (v/v) fetal calf serum (FCS) (Promocell, C-37350). Cells were kept under standard cell culture conditions (37 °C, 100% humidity, 5% CO2) and used at passage 2 to 4 for experiments.

The human colorectal cancer cell line Caco2 and human breast cancer cell lines Hs 578t, MCF-7, and MDA-MB-468 were obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Caco-2 cells were cultured in MEM with 20% FBS. Hs 578t cells were cultured in DMEM with 0.01 mg/mL insulin and 10% FBS. MCF-7 cells were cultured in MEM with 0.01 mg/mL insulin and 10% FBS. MDA-MB-468 cells were cultured in Leibovitz’s L-15 medium with 10% FBS. The human glioblastoma cell line U-87 MG, lung cancer cell Line A549, mouse teratoma cell line F9, and mouse colorectal cancer cell line CT26 were acquired from Procell Life Science Co., Ltd. (Wuhan, China) U87MG cells were cultured in MEM with 10% FBS. A549 cells were cultured in Ham’s F-12K medium with 10% FBS. F9 cells were cultured in DMEM with 10% FBS. CT26 cells were cultured in 1640 medium with 10% FBS. The HUVEC line was purchased from Promocell and cultured in endothelial cell growth medium (C-22110, Promocell, Heidelberg, Germany). 293F cells were a gift from Dr. Yang and were cultured in 293 expression medium (12338018, Thermo Fisher Scientific, Waltham, MA, USA). Each vial of cells was subjected to subculture for up to two weeks after recovery.