摘要
本文系统地研究了从人血白细胞膜（Buffy Coat,BC）中提取纯化单个核细胞（PBMC）的方法,以及以PBMC为原料批量生产高效价IL-2的一些条件。选择了既适合批量生产又能保证细胞质量的甲基纤维素-Ficoll-Hypaque（MC-FH）分离细胞法,分离的PBMC胎盘蓝排斥率高于95％,吞噬细胞含量低于5％。经过对各种诱生方法及诱生条件的研究,选择了TPA/PHA联合诱生方法;细胞浓度1.25～2.5×10<sup>6</sup>/ml为宜;以5％正常人AB血清（ABS）代替小牛血清（FCS）;用国产微生物多用培养箱代替进口CO<sub>2</sub>培养箱,在500ml血浆瓶中,37℃密闭旋转培养72～96小时。在以上条件下生产的IL-2效价一般可以达到4位数,最高为2100IU/ml,比PHA对照组诱生的IL-2最高效价提高20倍。一次从20单位白膜（来自4000ml全血）诱生IL-2产量为2～3×10<sub>6</sub>IU,可收获2000～3600ml诱生液。为利用白细胞膜原料生产IL-2提供了一个适合的生产工艺。
The methods of extraction of purified blood mononuclear cell(PBMC)from human bully coat (BC), and the conditions of serial production of high titer IL-2 by using PBMC as raw material were studied. For separation of PBMC was selected Methyl Cellulose-Ficoll Hypaque (MC-FH) which can not only be suited to its serial production but also ensure its quality. The separated PBMC's viability was more than 95%, and the phagocyte contamination was less than 5%. The combined PHA/TPA induction method was selected by investigating the various induction methods and conditions. The conditions for inducing the IL-2 by TPA/PHA method were as following., the PBMC's concentration had better be 1.25～2.5×10~6ml; 5% of normal human AB serum was substituted for calf serum; domestic universal microorganism culture kit was used instead of imported CO_2 culture kit: culture was conduced in the rotating 500ml sealed bottles at 37℃ for 72～96 hours. The titer of IL-2 produced under mentioned above conditions reached four digits, and its highest value was 2100 IU/ml and was 20 times higher than that of the titer of IL-2 induced by PHA method. The yield of IL-2 induced with 20units of buffy coat(from 4000ml whole blood) was 23×10~6IU, and the resulting amount of induced liquid was 2000～3600ml. This provided a suitable technological process for production of IL-2 using the buffy coat as raw meterial. (Original article on page 1)
出处
《中国输血杂志》
CAS
CSCD
1992年第1期1-5,共5页
Chinese Journal of Blood Transfusion
