In Vitro Culturing and Characteristics of Tumor-infiltrating Lymphocyte of Hepatocellular Carcinoma

The aim was to investigate the methods of ex vivo expansion of tumor-infiltrating lympho- cyte (TIL) obtained from hepatocellular carcinoma (HCC) biopsy specimens with a rapid expansion protocol. In this study, the specimen of primary liver cancer came from adjacent carcinoma tissues. After crushing and enzy- matic digestion, the lymphocytes in the single-cell slurry were separated by density centrifugation. The isolated lymphocytes were triggered with high dose of recombinant human interleukin 2 (IL-2) (2 000 U/mL), then were expanded in large-scale using peripheral blood allogeneic mononuclear cells as feeder cells. Nine primary HCC samples were used to try to isolate and culture the TIL in vitro. After two-step culturing, the number of TIL could achieve (1.0~5.5)×10 9 cells, and the expansion fold was 111 to 572. After expansion, the ratio of CD3+ T cells was (90.3±9.4)%, the ratio of CD3+CD4+ T cells was (24.9±14.1)%, the ratio of CD3+CD8+ was (56.4±20.2)%, and the ratio of CD3+CD56+ was (14.8±12.6)%. Moreover, the TIL showed strong killing activity to HepG2 and Bel-