Shimizu K, Park K, Yamada M, Tarura K, Matsui Y, Hayakawa T
Department of Neurosurgery, Osaka University Medical School, Japan.
J Clin Lab Immunol. 1990 May;32(1):41-7.
A high density cell culture system has been developed for large-scale production of lymphokine-activated killer (LAK) cells from peripheral blood lymphocytes (PBLs) of malignant tumor patients. The system consists of a culture bag, which has two compartments separated by a semipermeable membrane, and an external rotator. The system allows for a long-term, at least 4 weeks, culture of LAK cells at high cell density in the inner compartment. The collected PBLs were first divided between the two culture bags and cultured without harvesting for 7-10 days to obtain LAK cells. Half of the LAK cells from each bag was administered to patients twice a week for clinical trials. Culture of the remaining half was continued following addition of a fresh culture medium. LAK cells were transferred to patients alternatively from each bag for the following 2-3 weeks. The total number of LAK cells administered amounted to 3.9-9.8 (mean 5.8) times more than the PBLs collected by leukapheresis (n = 10). The 5 x 10(6)/ml of PBLS of the initial concentration reached a maximum of 2 x 10(7)/ml. Our system does not need for a CO2 incubator. Cytotoxicity of the LAK cells was evaluated in 4 hr 51Cr release assays. Mean cytotoxicity at maximum cell density was 95.4 +/- 3.2% against ONS-12 (a human glioma cell) and 84.8 +/- 3.0% against Daudi cells (n = 10), but gradually decreased to about 50% at the end of fourth week of the culture period. Cell viability of the LAK cells was normally over 80% through the entire culture period.(ABSTRACT TRUNCATED AT 250 WORDS)
已开发出一种高密度细胞培养系统，用于从恶性肿瘤患者的外周血淋巴细胞（PBL）大规模生产淋巴因子激活的杀伤细胞（LAK细胞）。该系统由一个培养袋和一个外部旋转器组成，培养袋有两个由半透膜隔开的隔室。该系统允许在内隔室以高细胞密度长期培养LAK细胞，培养时间至少为4周。收集的PBL首先在两个培养袋之间分配，培养7 - 10天不收获以获得LAK细胞。每个袋中一半的LAK细胞每周给患者给药两次用于临床试验。添加新鲜培养基后继续培养剩余的一半。在接下来的2 - 3周内，从每个袋中交替将LAK细胞转移给患者。给药的LAK细胞总数比通过白细胞分离术收集的PBL多3.9 - 9.8倍（平均5.8倍）（n = 10）。初始浓度为5×10⁶/ml的PBL最高达到2×10⁷/ml。我们的系统不需要二氧化碳培养箱。通过4小时⁵¹Cr释放试验评估LAK细胞的细胞毒性。在最大细胞密度下，对ONS - 12（一种人胶质瘤细胞）的平均细胞毒性为95.4±3.2%，对Daudi细胞的平均细胞毒性为84.8±3.0%（n = 10），但在培养期第四周结束时逐渐降至约50%。在整个培养期内，LAK细胞的细胞活力通常超过80%。（摘要截断于250字）
