Streptomycin

The human bladder cancer cell line T24 was a gift from the Molecular Tumour and Epigenetic Experiment of Chongqing Medical University (Chonqing, China). THP-1 human acute monocytic leukaemia cells were kindly provided by the Stem Cell Bank, Chinese Academy of Sciences. All cell lines were maintained in RPMI-1640 medium (cat. no. 10-040-CVR; Corning, Inc.) supplemented with 10% EV-depleted foetal bovine serum (FBS; cat. no. SA102.02; CellMax), 100 U/ml penicillin and 100 µg/ml streptomycin (cat. no. C0222; Beyotime Institute of Biotechnology), and bovine EV-depleted medium was obtained by overnight ultracentrifugation at 100,000 × g at 4°C (14 (link)). The cells were cultured at 37°C in a standard humidified incubator with 5% CO₂. All experiments were performed when the cells reached 70-80% confluency.

Human skin fibroblast cells (HSFs) were cultured on PS dishes in Dulbecco's modified Eagle's medium (DMEM; CellMax) supplemented with 10% fetal bovine serum (FBS, CellMax) and 1% antibiotic solution containing 10,000 units/mL penicillin, and 10,000 mg/mL streptomycin (CellMax). Then, the cells were incubated in a humidified atmosphere with 5% CO₂ at 37 °C. The cultured cells were harvested with 0.25% trypsin/EDTA (CellMax) and suspended in fresh culture media for the next subsequent subculture experiments. The same method was performed for the culture of human umbilical vein endothelial cells (HUVECs). HSFs and HUVECs were purchased from the American Type Culture Collection (ATCC, USA).