约氏疟原虫红外期体外培养及影响因素的研究

IN VITRO CULTIVATION OF THE EXOERYTHROCYTIC STAGE OF PLASMODIUM YOELII AND AFFECTING FACTORSINVOLVED

目的 :Wistar大鼠原代肝细胞 -约氏疟原虫红外期体外培养及若干因素的初步探讨。方法 :体外培养。结果 :用 15%小牛血清培养基培养原代肝细胞 ,培养密度为 2万 /孔 ,孔径为 16mm,接种约氏疟原虫子孢子后培养 4 8h,获得红外期疟原虫 ( EEF) ,其肝细胞感染率达 0 .0 35%±0 .0 13% ;接近成熟的 EEF直径达 4 0 .3× 31.6μm,有 10 0多个核 ,其 EEF可达 4 - 10个 / cm2 。经0 .0 1%乙二胺四乙酸 ( EDTA)洗脱肝细胞 ,离心 ,腹腔接种小鼠 ,第 7d血检原虫阳性。肝细胞密度为 0 .4万 /孔、 4万 /孔组及 10 %小牛血清组均未见 EEF。结论 :用含 15%小牛血清培养基、培养密度为 2万 /孔的 Wistar大鼠原代肝细胞较适合约氏疟原虫红外期的发育。

AIM:To cultivate the exoerythrocytic stage of Plasmodium yoelii in vitro and to study some involved affecting factors.METHOD:In vitro cultivation.RESULTS:The monolayer hepatocytes grown in 1 6 - mm plastic cell culture dishes were inoculated with sporozoite sus- pension prepared from Anopheles stephensi mosquitoes for48hours.At a final density of2× 1 0 4 cells per well,the infection rate of hepatocyte,cultured in medium supplemented wit15 % bovine serum,was 0 .0 35± 0 .0 1 3% ,the diameter of the nearly mature EEF of Plas- modium yoelii was up to40 .3× 31 .6 μm,and contained more than1 0 0 nuclei,the number of EEF might be4- 1 0 /cm2 .An intraperitoneal inoculation of the EE schizonts to mice could induce parasitemia.At a final density of0 .5× 1 0 4 or4× 1 0 4 cells per well or the hepatocytes cultured in medium supplemented with1 0 % bovine serum,no EEF could be observed.CON- CL USION:The density of hepatocytes and culture medium are important for the cultivation of the EE stage of Plasmodium yoelii.This procedure will lay foundation for the further studies of the sporozoite invasion,the development of EEF and the affecting factors in- volved.