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Updated: Jun 28, 2025
Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection
Published on: September 14, 2019
Cultivation, cryopreservation, and transcriptomic studies of host-adapted Cryptosporidium parvum and Cryptosporidium
Miner Deng1, Tianyi Hou1, Jie Zhang1
1State Key Laboratory for Animal Disease Control and Prevention, South China Agricultural University, Guangzhou, Guangdong, China.
Abstract:
Cryptosporidium hominis and Cryptosporidium parvum are major causes of severe diarrhea. Comparative studies of them are hampered by the lack of effective cultivation and cryopreservation methods, especially for C. hominis. Here, we describe adapted murine enteroids for the cultivation and complete development of host-adapted C. parvum and C. hominis subtypes, producing oocysts infectious to mice. Using the system, we developed a cryopreservation method for Cryptosporidium isolates. In comparative RNA-seq analyses of C. hominis cultures, the enteroid system generated significantly more host and pathogen responses than the conventional HCT-8 cell system. In particular, the infection was shown to upregulate PI3K-Akt, Ras, TNF, NF-κB, IL-17, MAPK, and innate immunity signaling pathways and downregulate host cell metabolism, and had significantly higher expression of parasite genes involved in oocyst formation. Therefore, the enteroid system provides a valuable tool for comparative studies of the biology of divergent Cryptosporidium species and isolates.
Insights
Researchers developed a novel murine enteroid system for cultivating Cryptosporidium parasites, enabling effective cryopreservation and comparative studies. This breakthrough aids in understanding Cryptosporidium hominis and Cryptosporidium parvum infections.
Area of Science:
- Microbiology
- Infectious Diseases
- Parasitology
Background:
- Cryptosporidium hominis and Cryptosporidium parvum are significant causes of severe diarrheal illness worldwide.
- Current research is limited by inadequate methods for cultivating and cryopreserving these parasites, particularly C. hominis.
Purpose of the Study:
- To establish a reliable method for cultivating and cryopreserving Cryptosporidium species.
- To facilitate comparative biological studies between C. hominis and C. parvum.
Main Methods:
- Adapted murine enteroids were utilized for the cultivation of host-adapted C. parvum and C. hominis subtypes.
- A novel cryopreservation technique for Cryptosporidium isolates was developed using the enteroid system.
- Comparative RNA-sequencing (RNA-seq) analyses were performed on C. hominis cultures.
Main Results:
- The murine enteroid system successfully supported the complete development of C. parvum and C. hominis, yielding infectious oocysts.
- The enteroid system demonstrated superior generation of host and pathogen responses compared to the conventional HCT-8 cell system.
- Infection upregulated key signaling pathways (PI3K-Akt, Ras, TNF, NF-κB, IL-17, MAPK) and innate immunity, while downregulating host cell metabolism. Enhanced expression of parasite genes related to oocyst formation was observed.
Conclusions:
- Murine enteroids provide a robust platform for cultivating and cryopreserving diverse Cryptosporidium species and isolates.
- This system significantly advances comparative research into the biology of C. hominis and C. parvum.
- The findings offer a valuable tool for future studies on Cryptosporidium pathogenesis and treatment strategies.
