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Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection
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JoVE 杂志 免疫与感染
Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection

Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection

DOI:
10.3791/59610-v

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05:31 min

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September 14, 2019

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, ,
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

Chapters

  • 00:04Title
  • 00:34In Vitro Sporozite Purification
  • 02:24Oocyst/Sporozoite Injection into the Organoid Lumen
  • 03:20Results: Representative Oocyst Identification and Analyses
  • 04:49Conclusion

Summary

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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

CryptosporidiumMicroinjectionSporozoite PurificationOocystExcystationMicroinjectorDMEMFast Green DyeL-glutathioneBetaineL-cysteineLinoleic AcidTaurineMicropipette PullerGlass Injection Capillaries

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