Esophageal Cell Line Cultivation and Characterization

Human ESCC KYSE-30, KYSE-150, KYSE-270, and KYSE-410 cell lines were obtained from Dr. C.S. Yang at Rutgers University and they are cultured in RPMI-1640/Ham’s F12 (1:1) medium with 5% FBS^{53 (link)}; EAC cell line OE33 (Sigma-Aldrich, USA) was cultured in RPMI-1640 medium containing 10% FBS; normal esophageal epithelial HET-1A cells (ATCC, USA) were maintained in serum-free LHC-9 medium^{53 (link)}. ATAC-seq analysis has been conducted in HET-1A and ESCC cell lines. The transcriptome profiling is consistent to published data. Cells are periodically examined to ensure free from mycoplasma contamination using Commercial Detection Kit (Lonza, Switzerland, LT07-703). One week before experiment, HET-1A cells were adapted in experimental culture medium same as that used for ESCC cells. All culture medium contained 1% penicillin/streptomycin and cells were maintained at 37 °C in a humidified 5% CO₂ incubator.