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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、繁殖
Adult Schistosoma japonicum cultured cells on the fifth day of culture were cultured in the presence of MNNG at final concentrations of 0 (control), 1, 2, 3, 4, 6, 9 μg / ml, % Calf serum and constant antibiotics in RPMI 1640 medium for 24 h, 36 h and 48 h, respectively, and then cultured in RPMI 1640 medium containing 20% calf serum and constant antibiotics. After 3 days, RPMI 1640 medium containing 5% calf serum and constant antibiotics was continued to culture and observed daily under an OlympusIM inverted microscope. Adult Schistosoma japonicum cultured cells treated with MNNG at a concentration of 6 μg / ml and 9 μg / ml were detached for about 3 days after treatment. A small amount of remaining cells were cultured in RPMI 16 40 medium supplemented with 5% fetal bovine serum and constant antibiotics Gradually died. The cultured cells of adult Schistosoma japonicum treated with MNNG at concentrations of 1, 2, 3 and 4μg / ml grew well. Compared with the control group, the cultured cells became larger and the number of dividing cells increased, especially at 3μ / ml MNNG 48h treatment group most obvious, and the observed foci, but subculture did not succeed. The results showed that to a certain extent, MNNG could induce foci of Schistosoma japonicum adult cells and induce fission and propagation