Three-dimensional (3D) culture of adult murine colon as an in vitro model of cryptosporidiosis: Proof of concept

Abstract

Cryptosporidium parvum is a major cause of diarrheal illness and was recently potentially associated with digestive carcinogenesis. Despite its impact on human health, Cryptosporidium pathogenesis remains poorly known, mainly due to the lack of a long-term culture method for this parasite. Thus, the aim of the present study was to develop a three-dimensional (3D) culture model from adult murine colon allowing biological investigations of the host-parasite interactions in an in vivo-like environment and, in particular, the development of parasite-induced neoplasia. Colonic explants were cultured and preserved ex vivo for 35 days and co-culturing was performed with C. parvum. Strikingly, the resulting system allowed the reproduction of neoplastic lesions in vitro at 27 days post-infection (PI), providing new evidence of the role of the parasite in the induction of carcinogenesis. This promising model could facilitate the study of host-pathogen interactions and the investigation of the process involved in Cryptosporidium-induced cell transformation.

Cellular immune response and scanning electron microscopy in the evaluation of Moringa leaves aqueous extract effect on Cryptosporidium parvum in buffalo intestinal tissue explants

“…Ileal tissue explants were prepared as previously described by Randall et al (2011) and Baydoun et al (2017) with some modifications. In brief, Ileal parts were cut into small pieces (40-60 mg) with a scalpel, then some of these pieces were placed into 24-well culture plates (one piece/ well) containing 1.5 ml RPMI 1640 medium with L-glutamine, supplemented with 1% Gentamicin 50 lg/ml and 10% FBS.…”

Metabolic Signatures of Cryptosporidium parvum-Infected HCT-8 Cells and Impact of Selected Metabolic Inhibitors on C. parvum Infection under Physioxia and Hyperoxia

“…Cryptosporidium parvum is one of the most important causal agents of human and animal cryptosporidiosis [ 1 , 2 , 3 , 4 , 5 ]. This zoonotic parasite (subphylum Apicomplexa) is considered the second-leading aetiology of diarrhoea-related mortality in children [ 4 , 6 , 7 ], and responsible for almost a million human deaths each year [ 8 , 9 ].…”

Phenotypic screening techniques forCryptosporidiumdrug discovery

“…Interestingly, although only immunocompromised mice are susceptible to C. parvum infection in vivo by oral gavage, the enteroids derived from immunocompetent adult mice were successfully infected with C. parvum sporozoites, although the authors noted inhibition of enteroid propagation after infection. A subsequent study from Baydoun et al looked at colonic explants from adult SCID mice and were able to successfully co-culture infected explants for 27 days (35 days uninfected) [120]. Yet another study looked at human intestinal and lung organoids derived from adult epithelial stem cells to support Cryptosporidium growth [121].…”