Culturing ESCC, HLEC, and NE-1 Cell Lines

ESCC cell lines KYSE30 (BW-S1092), KYSE150 (HT-X2475), EC109 (BW-5999), and EC9706 (BW-6316) were obtained from Cell Bank of Chinese Academy of Sciences (Shanghai, China) and kept in RPMI-1640 medium (#11875-093, Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS; #16000-044, Grand Island, NY, USA) at 37 °C in a 5% CO₂ cell culture incubator. Human lymphatic endothelial cells (HLECs, CP-H026, Procell) were obtained from ScienCell Research Laboratories (Carlsbad, CA, USA) and cultured in endothelial cell medium (ECM) with 5% FBS and endothelial cell growth medium supplements (CC-3121, Lonza, Basel, Switzerland). Immortalized esophageal epithelial NE-1 cell line (CP-H031, Procell) was obtained from State Key Laboratory of Oncology in South China, Sun Yat-sen University Cancer Center and cultured in Dulbecco’s modified Eagle medium (DMEM; #11885-076, Gibco, Grand Island, NY, USA) with 10% FBS in a 5% CO₂ atmosphere at 37 °C.