Cryopreservation of infectious Cryptosporidiumparvum oocysts

Cryptosporidiosis in an enteric infection caused by Cryptosporidium parasites and is a major cause of acute infant diarrhea in the developing world. A major bottleneck to research progress is the lack of methods to cryopreserve Cryptosporidium oocysts, thus requiring routine propagation in laboratory animals. Here, we report a method to cryopreserve C. parvum oocysts by ultra-fast cooling. Cryopreserved oocysts exhibit high viability and robust in vitro excystation, and are infectious to interferon-γ knockout mice. The course of the infection is comparable to what we observe with unfrozen oocysts. Oocyst viability and infectivity is not visibly changed after several weeks of cryogenic storage. Cryopreservation will facilitate the sharing of oocysts from well-characterized isolates and transgenic strains among different laboratories. Research on Cryptosporidium parasites, a major cause of acute infant diarrhea, is hampered by the short shelf life of oocysts, thus requiring routine propagation in laboratory animals. Here, the authors report a method to cryopreserve viable and infectious Cryptosporidium oocysts by ultra-fast cooling.