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. 1994 Sep;39(5):279–285. doi: 10.1007/BF01519979

Long-term serum/plasma-free culture of human cytotoxic T lymphocytes induced from peripheral blood mononuclear cells

Shu Qin Liu 1, Reiko Shiba 1, Berm Soo Kim 1, Kaoru Saijo 1, Tadao Ohno 1,✉
PMCID: PMC11038377  PMID: 7987858

Abstract

Tumor-specific human cytotoxic T lymphocytes (CTL) were induced by co-culturing peripheral blood mononuclear cells with X-ray-irradiated human lung squamous carcinoma cells, SQ-5, in the medium supplemented with interleukin(IL)-1, IL-2, IL-4 and IL-6, and 5% autologous plasma for 3 or 5 days. The CTL grew in serum/plasma-free medium containing these four interleukins and 0.5% bovine serum albumin for over a month and maintained kiling activity of target cells within 48 h at an effector/target ratio of 1.25. Their growth was essentially dependent on the target SQ-5 cells, which were renewed every 5 days. Under these conditions, IL-4 and IL-6 could be omitted. When anti-CD3 monoclonal antibody was added to the serum/plasma-free medium supplemented with IL-1 and IL-2, the target tumor cells were not required to maintain the specific killing activity of the CTL. A large number of CTL (1011) were obtained in 35 days.

Key words: Cytotoxic T lymphocyte, Serum-free culture, Interleukin, Killing activity, Adoptive immunotherapy

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