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Research on this parasite has been impeded by many technical limitations, including the lack of cryopreservation methods. While cryopreservation of <jats:italic>Cryptosporidium<\/jats:italic> oocysts by vitrification was recently achieved, the method is restricted to small sample volumes, thereby limiting widespread implementation of this procedure. Here, a second-generation method is described for cryopreservation of <jats:italic>C. parvum<\/jats:italic> oocysts by vitrification using custom high aspect ratio specimen containers, which enable a 100-fold increase in sample volume compared to previous methods. Oocysts cryopreserved using the described protocol exhibit high viability, maintain in vitro infectivity, and are infectious to interferon-gamma (IFN-\u03b3) knockout mice. Importantly, the course of the infection is comparable to that observed in mice infected with unfrozen oocysts. Vitrification of <jats:italic>C. parvum<\/jats:italic> oocysts in larger volumes will expedite progress of research by enabling the sharing of isolates among different laboratories and the standardization of clinical trials.<\/jats:p>","DOI":"10.1038\/s41598-020-68643-6","type":"journal-article","created":{"date-parts":[[2020,7,16]],"date-time":"2020-07-16T10:02:53Z","timestamp":1594893773000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Cryopreservation of infectious Cryptosporidium parvum oocysts achieved through vitrification using high aspect ratio specimen containers"],"prefix":"10.1038","volume":"10","author":[{"given":"Justyna J.","family":"Jaskiewicz","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Derin","family":"Sevenler","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anisa A.","family":"Swei","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanni","family":"Widmer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mehmet","family":"Toner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Saul","family":"Tzipori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rebecca D.","family":"Sandlin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,7,16]]},"reference":[{"key":"68643_CR1","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1016\/S0140-6736(13)60844-2","volume":"382","author":"KL Kotloff","year":"2013","unstructured":"Kotloff, K. 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