甲基硝基亚硝基胍诱导日本血吸虫成虫培养细胞增殖的研究

The Influence of MNNG on the Proliferation of Cultured Cell from Adult Schistosoma Japonicum

将培养第 5天的日本血吸虫成虫培养细胞在含甲基硝基亚硝基胍 (MNNG)终浓度分别为 0 (对照 )、1、2、3、4、6、9μg/ml的附加 2 0 %小牛血清及常量抗生素的RPMI 16 40培养基中分别处理 2 4h、36h、48h ;随后 ,换用含 2 0 %小牛血清及常量抗生素的RPMI 16 40培养基培养 ;3d后 ,改用含 5 %小牛血清及常量抗生素的RPMI 16 40培养基继续培养 ,每天在OlympusIM倒置显微镜下观察。用浓度为 6 μg/ml、9μg/ml的MNNG处理的日本血吸虫成虫培养细胞在处理后 3天左右大片脱落 ,残留的少量细胞在换用 5 %小牛血清及常量抗生素的RPMI 16 40培养后逐渐死亡 ;用 1、2、3、4μg/ml浓度MNNG处理后的日本血吸虫成虫培养细胞 ,生长良好 ,与对照组相比 ,培养细胞的体积变大 ,分裂细胞增多 ,尤以 3μ/ml的MNNG处理48h的试验组最为明显 ,并观察到转化灶 ,但传代培养没有成功。结果表明在一定程度上 ,MNNG能够诱导日本血吸虫成虫培养细胞出现转化灶 ,并发生分裂。

The function of MNNG on the proliferation of cultured cells from adult Schistosome were cultured in medium RPMI_1640 containing 20% calf serum and a moderate amount of antibiotics.Five days later,they were treated by MNNG with concentrations of 0,1,2,3,4,6 and 9μg/ml for 0,24,36 and 48 hours,respectively.Afterwards they were moved into the original medium.Three weeks later,the rate of calf serum in the medium was adjusted into to 5%.The proliferation and growth of the treated cells were observed by microscope everyday.Most of the cells treated by MMNG with concentrations of 6 and 9μg/ml fell off from the wall of culture flasks after 3 dyas,and the remained cells cultured in the medium containing 5% calf serum declined gradually.All the cells treated by MNNG of 1,2,3, and 4μg/ml grew well in the medium.The volume of the cells was larger and more cells in division were observed in the first groups than those in the control.especially in the cells treated by MNNG of the concentration of 3μg/ml for 48 hours.MNNG with the moderate concentration can induce the proliferation and division of cultured cells from adult Schistosoma japonicum.