摘要
目的建立微小隐孢子虫体外感染犬肾细胞(MDCK细胞)模型,并观察其生长发育过程。方法利用MDCK细胞为隐孢子虫感染对象,优化隐孢子虫感染MDCK细胞的培养条件,观察隐孢子虫在MDCK细胞中的生长发育过程。将体外感染48h的细胞培养上清接种小鼠,观察其感染情况。结果在含有5%胎牛血清的DMEM培养基中,用1×105隐孢子虫卵囊感染2.0×105个MDCK细胞,培养12h为最佳培养条件。在感染后72h内,隐孢子虫出现连续发育阶段,包括脱囊、子孢子、裂殖子、裂殖体、滋养体、配子体、合子、薄壁卵囊和厚壁卵囊,在60～72h内形成卵囊;用感染48h的细胞培养上清接种于免疫抑制小鼠,10d后有隐孢子虫卵囊排出。结论建立了能稳定用于微小隐孢子虫体外感染的MDCK细胞模型,观察到隐孢子虫的生长发育全过程。
Objective To develop an in vitro culture system for Cryptosporidium parvum in Madin-Darby canine kidney （MDCK） cell and observe its life cycle （from desquamate to oocyst）. Methods Oocysts of C. parvum were co-cultured with MDCK cells in vitro. Culture condition was optimized and the life cycle of C.parvum investigated. Results The optimal culture conditions for C. parvum in MDCK cells were 2.0×10^5 cells cultured for 12 h, and infected by 1.0×10^5 oocysts in the Dulbecco＇s Modified Eagle Medium with 5% FBS. Following 72 h co-culture, desquamate, sporozoites, trophozoites, meronts, microgametocytes, macrogametocytes, zygote, thin-wall oocyst, and thick-wall oocyst appeared orderly. Between the 60th and 72th hour, many oocysts emerged. Inoculated by the C. parvum-infected cell culture supernatant at the 48th hour, the immunosuppressed mice became infected. Conclusion The culture system provides a model for propagation of the parasites and demonstrates a complete in vitro life cycle of C. parvum.
出处
《中国寄生虫学与寄生虫病杂志》
CAS
CSCD
北大核心
2009年第3期219-222,共4页
Chinese Journal of Parasitology and Parasitic Diseases
基金
高等学校博士学科点专项科研基金(No.20030307022)
国家自然科学基金(No.39870575)~~
