Journal
/
Immunology and Infection
/
Studying Cryptosporidium Infection in 3D Tissue-derived Human Organoid Culture Systems by Microinjection
JoVE Journal
Immunology and Infection
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Immunology and Infection
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

•

05:31 min

•

September 14, 2019

•

, ,
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

Automatic Translation

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

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/59610?article_type=v

Assessing the Innate Sensing of HIV-1 Infected CD4+ T Cells by Plasmacytoid Dendritic Cells Using an Ex vivo Co-culture System.
check_url/59610?article_type=v

A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection
check_url/59610?article_type=v

Organoids as Model for Infectious Diseases: Culture of Human and Murine Stomach Organoids and Microinjection of Helicobacter Pylori
check_url/59610?article_type=v

Mouse- and Human-derived Primary Gastric Epithelial Monolayer Culture for the Study of Regeneration
check_url/59610?article_type=v

Quantification of Efferocytosis by Single-cell Fluorescence Microscopy
check_url/59610?article_type=v

Measuring Dengue Virus RNA in the Culture Supernatant of Infected Cells by Real-time Quantitative Polymerase Chain Reaction
check_url/59610?article_type=v

Analysis of Interactions between Endobiotics and Human Gut Microbiota Using In Vitro Bath Fermentation Systems
check_url/59610?article_type=v

Purification of High Yield Extracellular Vesicle Preparations Away from Virus
check_url/59610?article_type=v

Pneumococcus Infection of Primary Human Endothelial Cells in Constant Flow
check_url/59610?article_type=v

Measuring Phagocytosis of Aspergillus fumigatus Conidia by Human Leukocytes using Flow Cytometry

Read Article