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

Abstract: The aim was to investigate the methods of ex vivo expansion of tumor-infiltrating lymphocyte (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 enzymatic 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)×109 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 7402 cell lines in vitro. Taken together， the TIL from hepatocellular carcinoma can be successfully cultured and expanded in vitro， and with strong anti-tumor activity. It can be served as a means of the candidate immunotherapy for postoperative HCC patients.

CSTR: 32200.14.cjcb.2014.07.0017