›› 2009, Vol. 27 ›› Issue (3): 7-222.

• 实验研究 • Previous Articles     Next Articles

In vitro Cultivation Model of Cryptosporidium parvum in MDCK Cells and its Development

CHEN Fu1, 2, HUANG Ke-he1 *   

  1. 1 College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China；2 College of Animal Science and Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-06-30 Published:2009-06-30
  • Contact: HUANG Ke-he

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Abstract:

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 2.0×105 cells cultured for 12 h, and infected by 1.0×105 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.

Key words: Cryptosporidium parvum, Infection model, MDCK cell, Culture