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本文介绍我室有关LAK、TIL的体外诱导与活性调节研究及脐血CIK细胞诱导的初步研究.制备LAK-CM与PHA-CM(略)将TIL细胞(2×10~5/ml)分悬于含IL-2(500μ/ml)和/或PJ-CW(25μg/ml)的培液中置37℃,5%CO_2环境中培养,于培养后不同时间测定细胞扩增速度,并于第9天测定TIL细胞表型的变化(采用APAAP法与荧光检测法)及对自体肿瘤细胞与瘤靶细胞的杀伤活性(MIT显色法).将脐血淋巴细胞(1×10~6/ml)分别置含IL-2(2000μ/ml)和/或PJ-CW(25μg/ml)的环境中常规孵育,并于孵育后第5天进行表型分析(方法同前)及细胞毒活性测定.分离脐血淋巴细胞,于培养的不同时间分别加入rIFN-γ(1000U/ml),CD3MAB(20μl/ml),IL-2(300U/ml)及IL-1(100U/ml),测定其表型(方法同前).
This article describes the in vitro induction and activity regulation of LAK and TIL in our laboratory and the preliminary study on the induction of umbilical cord blood CIK cells. Preparation of LAK-CM and PHA-CM (omitted) Separate TIL cells (2×10~5/ml) The cells were cultured at 37°C and 5% CO 2 in culture medium containing IL-2 (500μ/ml) and/or PJ-CW (25μg/ml), and the rate of cell expansion was measured at different times after culture. 9-day determination of TIL cell phenotypic changes (using APAAP and fluorescence detection) and killing activity against autologous tumor cells and tumor target cells (MIT coloration method). Cord blood lymphocytes (1×10~6/ml ) Incubate routinely in an environment containing IL-2 (2000 μg/ml) and/or PJ-CW (25 μg/ml), and perform phenotypic analysis (method as before) and cytotoxicity assay on the 5th day after incubation. Isolation of umbilical cord blood lymphocytes, adding rIFN-γ (1000 U/ml), CD3MAB (20 μl/ml), IL-2 (300 U/ml), and IL-1 (100 U/ml) to culture at different times Phenotype (method with the former).