Kohgo Y, Kanisawa Y, Sakamaki S, Nojiri S, Ueno Y, Ito Y, Niitsu Y, Hosoi S, Sato S, Urushizaki I
Fourth Department of Internal Medicine, Sapporo Medical College.
Hum Cell. 1988 Mar;1(1):54-9.
Lymphokine activated killer (LAK) cells are generated by culture of lymphocytes with interleukin 2 (IL-2) in short term culture (3 to 5 days) and are used for adoptive immunotherapy for advanced cancer patients. The culture condition hitherto reported are essentially based on the rotating culture system, in which the maximum cell density was at 2 X 10(6) cell/ml and the cell recovery was usually less than 100%. The inability to induce LAK cells efficiently in vitro made the culturing of cells for therapy rather difficult and costly work because the mean infusion dose of LAK cells of one patient requires more than 1 X 10(10)/ml. We have therefore attempted to culture lymphocytes in 10 times higher concentration comparing with conventional methods. By using a new dialyzing culture system under continuous regulation of the amount of infused IL-2, nutrition medium, and pO2 and pCO2, we could culture cells at 2 X 10(7)/ml for more than 21 days and the resulted LAK cells showed a 100 times increase of activity on a per cell basis. By limiting dilution procedure, these killer cells mostly express T cell markers such as CD3 and CD8 but dose not express CD16.
淋巴因子激活的杀伤细胞（LAK细胞）是通过在短期培养（3至5天）中将淋巴细胞与白细胞介素2（IL-2）共同培养而产生的，并用于晚期癌症患者的过继性免疫治疗。迄今报道的培养条件基本上基于旋转培养系统，其中最大细胞密度为2×10⁶个细胞/毫升，细胞回收率通常低于100%。由于一名患者的LAK细胞平均输注剂量需要超过1×10¹⁰/毫升，因此无法在体外有效地诱导LAK细胞使得用于治疗的细胞培养成为一项相当困难且成本高昂的工作。因此，我们尝试以比传统方法高10倍的浓度培养淋巴细胞。通过使用一种新的透析培养系统，在持续调节输注的IL-2量、营养培养基以及pO₂和pCO₂的条件下，我们能够以2×10⁷/毫升的浓度培养细胞超过21天，并且所产生的LAK细胞在单个细胞基础上显示出活性增加了100倍。通过有限稀释法，这些杀伤细胞大多表达T细胞标志物如CD3和CD8，但不表达CD16。
