Two- and Three-Dimensional Bioengineered Human Intestinal Tissue Models for Cryptosporidium.

Area of Science:

• Parasitology
• Cell Biology
• Infectious Diseases

Background:

• Conventional cell cultures inadequately model Cryptosporidium infection's in vivo conditions and parasite life cycle.
• Previous 3D silk scaffold models using transformed cells supported infection but lacked primary human cell relevance.
• Human intestinal enteroids (HIEs) derived from Lgr5+ stem cells offer a more representative model of human intestinal epithelium.

Purpose of the Study:

• To develop and describe advanced in vitro models for studying Cryptosporidium infection.
• To utilize human intestinal enteroids (HIEs) for more accurate modeling of parasite-host interactions.
• To evaluate 2D and 3D HIE-derived models, co-cultured with myofibroblasts, for Cryptosporidium research.

Main Methods:

• Development of 3D silk scaffold-based models using transformed human intestinal epithelial cells (IECs).
• Isolation and culture of human intestinal enteroids (HIEs) containing Lgr5+ stem cells.
• Dissociation and culture of HIEs into 2D monolayers or 3D structures on silk scaffolds, co-cultured with human intestinal myofibroblasts.

Main Results:

• Transformed IEC models supported Cryptosporidium parvum infection for up to two weeks, completing the parasite life cycle.
• HIEs recapitulate native intestinal physiology, 3D architecture, and cellular diversity.
• HIE-derived models provide a promising platform for studying Cryptosporidium-host interactions and ex vivo intervention screening.

Conclusions:

• Human intestinal enteroid (HIE) models represent a significant advancement over traditional cell cultures for Cryptosporidium research.
• These HIE models, particularly in 3D configurations, offer a more physiologically relevant system for studying parasite biology.
• The described HIE-based models are valuable tools for future research into Cryptosporidium pathogenesis and therapeutic development.