应用二氧化碳培养箱连续培养大量恶性疟原虫配子体  

CONTINUOUS CULTIVATION OF A LARGE NUMBER OF PLASMODIUM FALCIPARUM GAMETOCYTES IN CARBON DIOXIDE INCUBATOR *

作　　者：许学年[1] 冯正[1] 杨柏林[1] 王聚君[1] 胡薇[1] 倪齐珍[1] 

机构地区：[1]中国预防医学科学院寄生虫病研究所

出　　处：《中国寄生虫学与寄生虫病杂志》1998年第5期376-379,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金

摘　　要：目的：为建立在二氧化碳（ＣＯ２）培养箱中连续培养大量恶性疟原虫（Ｐ．ｆ）配子体（Ｇ）的方法。方法：使用加入不同量ＮａＨＣＯ３的培养基，观察比较Ｇ的消长。结果：在培养第５ｄ（ｄ５）出现Ｇ，至ｄ１１－ｄ１３达到高峰；实验组与对照组Ｇ的峰值分别为１．８８％±０．６１％和１．２９％±０．４１％（Ｐ＜０．０５），表明实验组培养后期使用含较高浓度ＮａＨＣＯ３的完全培养基，有助于Ｇ的分化。Ⅰ－Ⅴ期Ｇ的高峰分别出现于培养ｄ５、ｄ７、ｄ１１、ｄ１３和ｄ１５。ｄ１５Ｖ期Ｇ率为７．１％—５２．６％，平均为２４．３％。雌、雄Ｇ比例为１２．８∶１。原虫从液氮中取出复苏后，加入新鲜红细胞连续分皿２４代，仍保持较高的产生Ｇ的能力。经薄膜饲血器多次人工感染斯氏按蚊（Ａｎｏｐｈｅｌｅｓｓｔｅｐｈｅｎｓｉ），解剖３２８只蚊胃，未见卵囊。结论：本虫株使用该方法可以持续稳定地取得大量Ｐ．ｆＧ。AIM: To establish a method for continuously cultivating a large number of P. falciparum gametocytes in vitro using carbon dioxide incubator.METHODS: The number of gametocytes produced in experimental and control groups was compared after the addition of various concentrations of NaHCO 3 to the culture medium. RESULTS:The gametocytes began to rise on day 5 of cultivation and reached a peak on day 11-13. The peak gametocyte loads were 1.9%±0.6% and 1.3%±0.4%( P <0.05) in experimental and control group,respectively, indicating that the complete medium with 30 mmol/L sodium bicarbonate was benificial to gametogenesis. The numbers of stages Ⅰ-Ⅴ gametocytes rose to a peak on d5, d7, d11, d13 and d15, respectively. On day 15 the percentage of stage Ⅴ gametocytes was 7.1%-52.6% with an average of 24.3%. The ratio of macrogametocyte to microgametocyte was 12.8∶1. The parasites were able to produce high gametocytaemia up to 24th subculture after thawing. Laboratory reared Anopheles stephensi fed through membrane on blood infected with P.falciparum were disected, no oocyst was found in the midgut. CONCLUSION: A culture system which could consistently and stably produce a large number of gametocytes of P.falciparum was established.

关 键 词：恶性疟原虫 配子体 培养箱 二氧化碳 

分 类 号：R382.31[医药卫生—医学寄生虫学]