Murata M, Yano T, Togami M, Yasumoto K, Sugimachi K, Kimura G, Nomoto K
Department of Immunology, Kyushu University, Fukuoka, Japan.
J Immunol Methods. 1990 May 8;129(1):89-95. doi: 10.1016/0022-1759(90)90424-t.
We have developed a new culture system (termed the JCC device) for the generation and expansion of human lymphokine-activated killer (LAK) cells. The JCC device was essentially based on dialysis culture. A dialysis membrane divided the culture vessel from the outside circulation of basal medium. Human lymph node lymphocytes were cultured in the culture vessel using a culture volume of 100 ml at an initial cell density of 1 X 10(6)/ml in the presence of recombinant human interleukin-2. The culture vessel was rotated for the proper dispersion of cells. The cell density reached up to 1.2 X 10(7)/ml on day 17 of culture. A modified JCC device supplying oxygen through a silicon tube prevented oxygen depletion in the culture vessel. With this modified JCC device cell densities up to 4.3 X 10(7)/ml were achieved without reduction of cytolytic activity. Cell viability was more than 90% throughout the culture. The use of this JCC device permitted the culturing of LAK cells at a high cell density. The procedure is relatively cheap, simple and less time consuming than previous methods.
我们开发了一种新的培养系统（称为JCC装置），用于人淋巴因子激活的杀伤（LAK）细胞的生成和扩增。JCC装置主要基于透析培养。透析膜将培养容器与基础培养基的外部循环隔开。人淋巴结淋巴细胞在培养容器中培养，培养体积为100 ml，初始细胞密度为1×10⁶/ml，同时存在重组人白细胞介素-2。培养容器旋转以适当分散细胞。培养第17天时，细胞密度达到1.2×10⁷/ml。一种通过硅管供应氧气的改良JCC装置可防止培养容器中的氧气耗尽。使用这种改良的JCC装置，细胞密度可达4.3×10⁷/ml，而细胞溶解活性不会降低。在整个培养过程中，细胞活力超过90%。使用这种JCC装置可以在高细胞密度下培养LAK细胞。该方法相对便宜、简单，且比以前的方法耗时更少。
