Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by MicroinjectionDevanjali Dutta 1, Inha Heo 1,3, Roberta O'Connor 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
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.