A Cell Culture Platform for the Cultivation of Cryptosporidium parvum

Cryptosporidium is a genus of ubiquitous unicellular parasites belonging to the phylum Apicomplexa. Cryptosporidium species are the second largest cause of childhood diarrhoea and are associated with increased morbidity. Accompanying this is the low availability of treatment and lack of vaccines. The major barrier to developing effective treatment is the lack of reliable in vitro culture methods. Recently, our lab has successfully cultivated C. parvum in the oesophageal cancer derived cell line COLO-680N, and can maintain infection for several weeks. The success of this cell line was assessed with a combination of various techniques including fluorescent microscopy and qPCR. In addition, to tackle the issue of long-term oocyst production in vitro, a simple, low cost bioreactor system using the COLO-680N cell line was established, which produced infectious oocysts for four months. This chapter provides details on the methodologies used to culture, maintain and assess Cryptosporidium infection and propagation in COLO-680N.