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Immunology and Infection

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JoVE Journal Immunology and Infection
Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection
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Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection

Article DOI: 10.3791/59610-v • 05:31 min • September 14th, 2019
September 14th, 2019 •
1, 1,3, 2
1Hubrecht Institute, Oncode Institute, Royal Netherlands Academy of Arts and Sciences (KNAW), UMC Utrecht, 2Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, 3Janssen Pharmaceutica

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Summary
Automatic Translation

We describe protocols to prepare oocysts and purify sporozoites for studying infection of human intestinal and airway organoids by Cryptosporidium parvum. We demonstrate the procedures for microinjection of parasites into the intestinal organoid lumen and immunostaining of organoids. Finally, we describe the isolation of generated oocysts from the organoids.

Tags

Cryptosporidium Infection 3D Tissue-derived Human Organoid Culture Systems Microinjection Sporozoite Purification Oocyst Excystation Medium Wash Solution Centrifugation DMEM Concentration Filter Holder Apparatus Pore Size Polycarbonate Filter Sporozoite Suspension
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