Culturing Human Esophageal and Kidney Cell Lines

Human ESCC cell lines KYSE30, KYSE150, KYSE450, KYSE510, and TE-1 purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China) were cultured in RPMI 1640 medium (Gibco, USA) containing 10% fetal bovine serum (FBS; Gibco, USA) as previously described [42 (link)]. The immortalized human esophageal epithelial cell line Het-1A and human embryonic kidney 293T cells, obtained from the ATCC (Manassas, VA, USA), were cultured in Dulbecco’s modified Eagle’s medium culture medium supplemented with 10% FBS. Cells were kept in a humidified incubator with 5% CO₂ at 37 °C. All cells were periodically authenticated by using the short tandem repeat analysis and tested for mycoplasma contamination by PCR.