A High-Throughput Organoid Microinjection Platform to Study Gastrointestinal Microbiota and Luminal Physiology
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Williamson, Ian A, et al. A High-throughput Organoid Microinjection Platform to Study Gastrointestinal Microbiota and Luminal Physiology. 2018. https://doi.org/10.17615/ksb4-1h46APA
Williamson, I., Arnold, J., Samsa, L., Gaynor, L., Di Salvo, M., Cocchiaro, J., Carroll, I., Azcarate Peril, M., Rawls, J., Allbritton, N., & Magnuss, S. (2018). A High-Throughput Organoid Microinjection Platform to Study Gastrointestinal Microbiota and Luminal Physiology. https://doi.org/10.17615/ksb4-1h46Chicago
Williamson, Ian A., Jason W Arnold, Leigh Ann Samsa, Liam Gaynor, Matthew Di Salvo, Jordan L Cocchiaro, Ian Carroll et al. 2018. A High-Throughput Organoid Microinjection Platform to Study Gastrointestinal Microbiota and Luminal Physiology. https://doi.org/10.17615/ksb4-1h46- Creator
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Williamson, Ian A.
- Affiliation: School of Medicine, UNC/NCSU Joint Department of Biomedical Engineering
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Arnold, Jason W.
- Affiliation: School of Medicine, Department of Medicine
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Samsa, Leigh Ann
- Affiliation: School of Medicine, Joint Department of Biomedical Engineering, North Carolina State University
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Gaynor, Liam
- Other Affiliation: Harvard Medical School
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DiSalvo, Matthew
- Affiliation: School of Medicine, Joint Department of Biomedical Engineering, North Carolina State University
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Cocchiaro, Jordan L.
- Other Affiliation: Duke University
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Carroll, Ian
- Affiliation: School of Medicine, Department of Medicine
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Azcarate-Peril, M. Andrea
- Affiliation: School of Medicine, Department of Medicine
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Rawls, John F.
- Other Affiliation: Duke University
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Allbritton, Nancy L.
- Affiliation: School of Medicine, Joint Department of Biomedical Engineering, North Carolina State University
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Magnuss, Scott T.
- Affiliation: School of Medicine, Joint Department of Biomedical Engineering, North Carolina State University
- Abstract
- Background & Aims The human gut microbiota is becoming increasingly recognized as a key factor in homeostasis and disease. The lack of physiologically relevant in vitro models to investigate host–microbe interactions is considered a substantial bottleneck for microbiota research. Organoids represent an attractive model system because they are derived from primary tissues and embody key properties of the native gut lumen; however, access to the organoid lumen for experimental perturbation is challenging. Here, we report the development and validation of a high-throughput organoid microinjection system for cargo delivery to the organoid lumen and high-content sampling. Methods A microinjection platform was engineered using off-the-shelf and 3-dimensional printed components. Microinjection needles were modified for vertical trajectories and reproducible injection volumes. Computer vision (CVis) and microfabricated CellRaft Arrays (Cell Microsystems, Research Triangle Park, NC) were used to increase throughput and enable high-content sampling of mock bacterial communities. Modeling preformed using the COMSOL Multiphysics platform predicted a hypoxic luminal environment that was functionally validated by transplantation of fecal-derived microbial communities and monocultures of a nonsporulating anaerobe. Results CVis identified and logged locations of organoids suitable for injection. Reproducible loads of 0.2 nL could be microinjected into the organoid lumen at approximately 90 organoids/h. CVis analyzed and confirmed retention of injected cargos in approximately 500 organoids over 18 hours and showed the requirement to normalize for organoid growth for accurate assessment of barrier function. CVis analyzed growth dynamics of a mock community of green fluorescent protein– or Discosoma sp. red fluorescent protein-expressing bacteria, which grew within the organoid lumen even in the presence of antibiotics to control media contamination. Complex microbiota communities from fecal samples survived and grew in the colonoid lumen without appreciable changes in complexity. Conclusions High-throughput microinjection into organoids represents a next-generation in vitro approach to investigate gastrointestinal luminal physiology and the gastrointestinal microbiota.
- Date of publication
- May 2018
- Keyword
- DOI
- Identifier
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- Resource type
- Article
- Rights statement
- In Copyright
- Journal title
- Cellular and Molecular Gastroenterology and Hepatology
- Journal volume
- 6
- Journal issue
- 3
- Page start
- 301
- Page end
- 319
- Language
- English
- Version
- Postprint
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