A cell culture platform for Cryptosporidium that enables long-term cultivation and new tools for the systematic investigation of its biology

Abstract

Cryptosporidium parasites are a major cause of diarrhoea that pose a particular threat to children in developing areas and immunocompromised individuals. Curative therapies and vaccines are lacking, mainly due to lack of a long-term culturing system of this parasite. Here, we show that COLO-680N cells infected with two different Cryptosporidium parvum strains produce sufficient infectious oocysts to infect subsequent cultures, showing a substantial fold increase in production, depending on the experiment, over the most optimistic HCT-8 models. Oocyst identity was confirmed using a variety of microscopic- and molecular-based methods. This culturing system will accelerate research on Cryptosporidium and the development of anti-Cryptosporidium drugs.

Keywords: Atomic force microscopy; COLO-680N; Cell culture; Cryptosporidium; Electron microscopy; Immunofluorescence microscopy; Lipidomics; Proteomics.