探究获得高成熟率伯氏疟原虫裂殖体的体外培养条件∗

摘要

In order to obtain abundant and viable mature schizonts of Plasmodium berghei, many in vitro culture conditions of the P. berghei ANKA were optimized, such as the volume of culture medium, the percentage of fetal bovine serum in culture medium, the cell concentration of cultivation. In this study, when the parasitemia had reached to 1%–3%, most of the intra⁃erythrocytic stage parasites were ring or young trophozoites. After 18 to 24 hours in vitro cultivation, the infected blood was collected and purified to observe the percentage and viability of mature schizonts in different culture conditions. Compared with previous methods, P. berghei schizonts cultured with optimized culture condition can improve percentage of mature schizonts to 80%, and there are 12 -16 merozoits in one mature schizont. With schizonts re⁃invasion of new erythrocytes, the parasitemia was 1�57% by tail intravenous inoculation 4 hours later and increased 3�4 times compared to the control group. So the optimized culture condition can improve the yield and viability of schizonts which lay the foundation for transfection in P. berghei.%为获得大量有活力的伯氏疟原虫裂殖体，本研究对伯氏疟原虫ANKA虫株的体外培养条件主要从培养基的用量、培养基胎牛血清的含量、培养的细胞浓度、培养时间及培养的气体环境等方面进行了优化。当小鼠体内虫血率达到1％～3％，在红内期疟原虫处于环期或早期滋养体阶段时取血分组培养，观察在不同培养条件下裂殖体的状态并检测其活力。与既往的体外培养方法相比，优化后的培养方法，可将裂殖体的成熟率提高到80％，每个裂殖体含12～16个裂殖子，裂殖体尾静脉注射重新入侵红细胞4 h后的虫血率为1�57％，与对照组相比裂殖体的活力提高3�4倍。优化的方法可以提高裂殖体的得量和活力，为伯氏疟原虫的转染奠定基础。