Smmc 7721

We collected 40 pairs of liver tumors and adjacent tissues from patients treated at The First Affiliated Hospital of Chongqing Medical University. Patients were excluded if they receive chemotherapy or radiation before surgery. Our study was approved by the Ethics Committee of Chongqing Medical University and that this was conducted in accordance with the Declaration of Helsinki. We obtained written informed consent from each patient after providing information about the study, and patients signed the informed consent forms. The normal hepatocyte cell line (HL-7702) and human HCC lines (SMMC-7721, Hep-3B, Hep-G2, SMMC-7402, and Huh-7) were purchased from the Institute of Chinese Academy of Sciences. SMMC-7721, Hep-3B, Hep-G2, SMMC-7402 and Huh-7 cell lines were cultured in DMEM (Corning Incorporated, Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Thermo Scientific, Waltham, MA, USA). The HL-7702 cell line was cultured in RPMI 1640 medium (Corning Incorporated, Corning, NY, USA) supplemented with 10% FBS. All cells were cultured in an incubator with an atmosphere of 95% O₂ and 5% CO₂ at 37°C.

All the human HCC-derived cell lines were obtained from the American Type Culture Collection (ATCC), with exception of SMMC7721, which was obtained from China Infrastructure of Cell Line Resource. Huh7, Sk-hep-1, Hep3B, HepG2, PLC/PRF/5 and SMMC7721 were cultured in Dulbecco's modified Eagle's medium (DMEM) (Corning,10–013-CVR) while SNU387, SNU182, SNU423 and SNU475 were cultured in RPMI1640(Corning,10-040-CVR), both of which were supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. Cells were maintained at 37°C in a humid atmosphere containing 5% CO₂. Cell culture supernatants were collected from 1×10⁷ cells and added into PBMCs cultural system at a volume ratio of 1:4 and cultured overnight.