Chen Fu, Huang Ke-He
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2009 Jun;27(3):219-22.
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).
Oocysts of C. parvum were co-cultured with MDCK cells in vitro. Culture condition was optimized and the life cycle of C.parvum investigated.
The optimal culture conditions for C. parvum in MDCK cells were 2.0 x 10(5) cells cultured for 12 h, and infected by 1.0 x 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 72nd hour, many oocysts emerged. Inoculated by the C. parvum-infected cell culture supernatant at the 48th hour, the immunosuppressed mice became infected.
The culture system provides a model for propagation of the parasites and demonstrates a complete in vitro life cycle of C. parvum.
建立微小隐孢子虫在犬肾传代细胞（MDCK）中的体外培养体系，并观察其生命周期（从脱囊到卵囊形成）。
微小隐孢子虫卵囊与MDCK细胞进行体外共培养。优化培养条件并研究微小隐孢子虫的生命周期。
微小隐孢子虫在MDCK细胞中的最佳培养条件为：2.0×10⁵个细胞培养12小时，然后在含5%胎牛血清的杜氏改良 Eagle培养基中接种1.0×10⁵个卵囊。共培养72小时后，依次出现脱囊、子孢子、滋养体、裂殖体、小配子体、大配子体、合子、薄壁卵囊和厚壁卵囊。在第60至72小时之间，出现许多卵囊。用第48小时感染微小隐孢子虫的细胞培养上清液接种免疫抑制小鼠，小鼠被感染。
该培养体系为寄生虫繁殖提供了模型，并展示了微小隐孢子虫完整的体外生命周期。
