Kyse450

Six OSCC cell lines, WHCO1, WHCO3, WHCO5 (Veale and Thornley, 1984 ), Kyse30, Kyse150, and Kyse450 (obtained commercially from Deutsche Sammlung von Mikroorganismen und Zellkulturen), originally established from surgical specimens of primary OSCCs, were grown in DMEM (Life Technologies, Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS), 50 U of penicillin/ml, and 50 mg streptomycin/ml (Life Technologies, Carlsbad, CA). Primary normal esophageal epithelial cells (EPC2) established from normal human esophagus (Andl et al., 2003 (link)) were grown in keratinocyte-SFM medium (Life Technologies, Rockville, MD) containing bovine pituitary extract (40 mg/ml) (Life Technologies, Rockville, MD). All cells were maintained in a 5% CO₂ humidified incubator at 37ºC. Cell lines were routinely frozen within two passages after receipt from the cell bank and used within less than 6 mo after resuscitation.

The ESCC cell lines (KYSE150, KYSE410, KYSE70, KYSE510, KYSE450, EC1, TE7) were obtained from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ, Braunschweig, Germany). TE1 cells were purchased from the Cell Bank of the Chinese Academy (Shanghai, China). All ESCC cells were cultured in RPMI-1640 supplemented with 10% FBS and 100IU/ml Penicillin-Streptomycin solution at 37 °C in 5% CO2. TE1R and KYSE150R were previously established in our laboratory [28 (link)].
Twenty-four pairs of ESCC tumor samples and adjacent nontumor tissues were collected from patients who had undergone surgery and were histologically diagnosed at the First Affiliated Hospital of the Zhejiang University of China.

Normal esophageal epithelial cells (SHEE10) were obtained from The Cell Bank of Type Culture Collection of The Chinese Academy of Sciences, and four human ESCC cell lines (KYSE30, KYSE150, KYSE450 and KYSE510) were obtained from DSMZ-German Collection of Microorganisms and Cell Cultures GmbH. All cells were cultured in RPMI-1640 medium containing 10% FBS (both Thermo Fisher Scientific, Inc.), streptomycin (100 µg/ml) and penicillin (100 IU/ml) at 37°C with 5% CO₂.

One of the esophageal squamous cell carcinoma (ESCC) cell lines of Hong Kong Chinese origin, SLMT-1 [21 (link)], was kindly provided by Professor Gopesh Srivastava of the Department of Pathology, the University of Hong Kong. The other two ESCC cell lines of Hong Kong Chinese origin, including HKESC-2 and HKESC-4, and three ESCC cell lines of Japanese origin, including KYSE-150, KYSE-450, and KYSE-520 [22 (link)], were purchased from DSMZ (Braunschweig, Germany). Two non-tumor esophageal epithelial cell lines including NE-1 [23 (link)] and NE-3 [24 (link)] were kindly provided by Professor George S. W. Tsao from the Department of Anatomy, the University of Hong Kong. A human skin cell line, HEK001, was purchased from the American Type Culture Collection (ATCC) and cultured as suggested. The culture medium for KYSE-150 and KYSE-450 was RPMI with 45% F-12 and 10% FBS, that for KYSE-520 was RPMI with 10% FBS, that for SLMT-1, HKESC-2, and HKESC-4 was MEMα with 10% FBS, and NE-1, NE-3, and HEK001 were cultured on KSFM with complementary supplements. All media were supplemented with 100 units/mL penicillin G and 100 μg/mL streptomycin, and all cell lines mentioned above were cultured at 37°C in a humidified incubator with 5% CO₂.