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A Millifluidic Perfusion Cassette for Studying the Pathogenesis of Enteric Infections Using Ex-Vivo Organoids

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Abstract

To generate physiologically-relevant experimental models, the study of enteric diarrheal diseases is turning increasingly to advanced in vitro models that combine ex vivo, stem cell-derived “organoid” cell lines with bioengineered culture environments that expose them to mechanical stimuli, such as fluid flow. However, such approaches require considerable technical expertise with both microfabrication and organoid culture, and are, therefore, inaccessible to many researchers. For this reason, we have developed a perfusion system that is simple to fabricate, operate, and maintain. Its dimensions approximate the volume and cell culture area of traditional 96-well plates and allow the incorporation of fastidious primary, stem cell-derived cell lines with only minimal adaptation of their established culture techniques. We show that infections with enteroaggregative E. coli and norovirus, common causes of infectious diarrhea, in the system display important differences from static models, and in some ways better recreate the pathophysiology of in vivo infections. Furthermore, commensal strains of bacteria can be added alongside the pathogens to simulate the effects of a host microbiome on the infectious process. For these reasons, we believe that this perfusion system is a powerful, yet easily accessible tool for studying host-pathogen interactions in the human intestine.

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Acknowledgments

This work was supported in part by grants from the National Institutes of Health (U19 AI116497 and F30 DK108541) and CPRIT RP160283 – Baylor College of Medicine Comprehensive Cancer Training Program. The authors thank Drs. Noah Shroyer and Sue Crawford for helpful discussions and insights, Dr. Jim Broughman and Xi-Lei Zheng for technical assistance with the flow experiments, and Dr. Jennifer Connell for editorial assistance.

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Author notes

  1. Sarah A. Hewes and Anubama Rajan contributed equally .

Authors and Affiliations

  1. Department of Bioengineering, Rice University, 6100 Main St, MS-142, Houston, TX, 77005, USA

    Reid L. Wilson, Sarah A. Hewes, Takanori Iida & K. Jane Grande-Allen

  2. Department of Molecular Virology and Microbiology, Baylor College of Medicine, One Baylor Plaza, Houston, TX, 77030, USA

    Anubama Rajan, Shih-Ching Lin, Carolyn Bomidi, Mary K. Estes & Anthony W. Maresso

  3. Medical Scientist Training Program, Baylor College of Medicine, One Baylor Plaza, Houston, TX, 77030, USA

    Reid L. Wilson

  4. Department of Medicine, Baylor College of Medicine, One Baylor Plaza, Houston, TX, 77030, USA

    Mary K. Estes

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Correspondence to K. Jane Grande-Allen.

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Associate Editor Sriram Neelamegham oversaw the review of this article.

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Wilson, R.L., Hewes, S.A., Rajan, A. et al. A Millifluidic Perfusion Cassette for Studying the Pathogenesis of Enteric Infections Using Ex-Vivo Organoids. Ann Biomed Eng 49, 1233–1244 (2021). https://doi.org/10.1007/s10439-020-02705-8

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