【摘要】 目的 为了使多房棘球蚴在非寄生离体的情况下保持活力，利于移植，需要多房棘球蚴(Echinococcus multilocularis,Em)在体外不同影响因素下的最佳培养条件。方法 选择不同的培养条件：生理盐水培养组、无钙镁PBS培养组、血清培养组、无血清培养组，在两种温度(37℃,4℃)下培养多房棘球蚴，每组1×10~4个虫体，连续培养5 d,以经典的染色方法：台盼蓝染色法、亚甲基蓝染色法、伊红染色法、Dil染色法对虫株进行活力染色并计数。第一组小鼠注射新鲜离体虫株、第二组小鼠注射4℃生理盐水处理24 h的虫株，一月后观察两处理组小鼠肝脏变化。结果 随培养时间延长，台盼蓝染色结果显示，第1～5 d离体虫株生存率平均值分别为86.575%、94.600%、63.387%、51.225%、32.775%。差异有统计学意义(P<0.05)。生理盐水组、无钙镁PBS组、血清组、无血清组5 d体外培养平均生存率为50.95%、64.78%、72.50%、74.62%。台盼蓝染色结果显示，无血清组在4℃、37℃时平均生存率分别为56.475%、87.995%,两组间有统计学差异(P<0.05)。无血清组四种染色方法间无统计学差异(P>0.05)。离体虫株4℃静置24 h,生理盐水混悬注射组较新鲜离体虫株生理盐水混悬注射组小鼠动物造模感染程度低。结论 37℃无血清培养的多房棘球蚴绦虫，较其余三组培养条件的体外生存率更高。进行多房棘球蚴体外移植时效果更佳，为实验室提高泡球蚴动物造模成功率，延长虫体体外生存周期奠定了基础。更多还原

【Abstract】 Objective In order to maintain the vitality of Echinococcus multilocularis(Em) in non parasitic ex vivo conditions and facilitate transplantation, the optimal culture conditions for Em under different influencing factors in vitro are required. Methods Select different culture conditions: physiological saline culture group, calcium magnesium free PBS culture group, serum culture group, and serum free culture group. Cultivate multiple hydatid cells at two different temperatures(37 ℃,4 ℃),with 1×10~4 cells in each group for 5 consecutive days. Use classic staining Methods such as trypan blue staining, methylene blue staining, eosin staining, and Dil staining to stain and count the viability of the cells. The first group of mice were injected with fresh isolated worm strains, while the second group of mice were injected with worm strains treated with 4 ℃ physiological saline for 24 hours. One month later, the liver changes of the two treatment groups of mice were observed. Results With the extension of cultivation time, the results of trypan blue staining showed that the average survival rates of isolated insect strains from the first day to the fifth day were 86.575%,94.600%,63.387%,51.225%,and 32.775%,respectively. The difference was statistically significant(P<0.05). The average 5-day in vitro survival rates of the physiological saline group, calcium magnesium free PBS group, serum group, and serum free group were 50.95%,64.78%,72.50%,and 74.62%,respectively. The results of trypan blue staining showed that the average survival rates of the serum-free group at 4 ℃ and 37 ℃ were 56.475% and 87.995%,respectively, with statistical differences between the two groups(P<0.05). There was no statistically significant difference(P>0.05) among the four staining methods in the serum-free group. The isolated strain was left to stand at 4 ℃ for 24 hours, and the infection level in the physiological saline suspension injection group was lower than that in the fresh isolated strain physiological saline suspension injection group. Conclusion The in vitro survival rate of Em cultured at 37 ℃ without serum was higher than that of the other three groups of culture conditions. The effect of in vitro transplantation of Em is better, which lays the foundation for improving the success rate of animal modeling of AE in the laboratory and prolonging the in vitro survival cycle of the parasite.更多还原