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Organoids as a Model for Intestinal Ion Transport Physiology

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Ion Transport Across Epithelial Tissues and Disease

Part of the book series: Physiology in Health and Disease ((PIHD))

Abstract

The advent of intestinal organoid culture in 2009 was a fortuitous development in the search for a valid marker of intestinal stem cells, and provided proof of murine intestinal stem cell regenerative potential. Intestinal organoid culture was preceded by key discoveries of the Wnt/β-catenin signaling pathway and the development of 3D culture matrices. The latter, involving a laminin-rich gel to provide an artificial basement membrane, was instrumental to primary intestinal epithelial culture by preventing anoikis, an immediate apoptotic event when intestinal epithelial cells detach from the basement membrane. One of the first physiological studies using 3D murine “mini-gut” structures showed cystic fibrosis transmembrane conductance regulator (CFTR) expression and anion channel activity in the crypt-like structures projecting from the epithelial-lined central cavity. Detailed investigations of ion transport physiology using human intestinal organoids, both primary and iPSC-derived, found close similarities to existing knowledge of ion transport physiology and included the development of the forskolin-induced swelling assay (FIS). The FIS assay using organoids cultured from rectal biopsies of cystic fibrosis patients provided an avenue for personalized medicine to test small-molecule modulators on different CFTR mutations. More recent research has led to the development of 2D primary intestinal epithelial monolayers, which provide easy access to the apical, lumen-facing membrane and the opportunity for traditional ion transport studies with Ussing chambers. Human 2D primary intestinal monolayers also demonstrate the dominance of CFTR in anion secretion and provide a quantitative evaluation of its chloride and bicarbonate secretory conductances. These aspects of ion transport physiology using 2D and 3D intestinal cultures are discussed along with the relative advantages and disadvantages of each culture method with respect to technical aspects and recapitulation of native intestinal epithelium.

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Acknowledgments

The authors would like to thank Drs. Rowena Woode and Sarah Young, University of Missouri, for thoughtful review and comments on the manuscript. Supported by grants NIH R01DK048816 (LLC); Cystic Fibrosis Foundation grants CLARKE16P0, CLARKE17G0, CLARKE19XX0, DEJONG19GO, CF Foundation Therapeutics (HRdJ) and SRC011, UK-CF Trust (HRdJ).

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Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, Erasmus MC University Medical Center, Rotterdam, The Netherlands

    Hugo R. de Jonge & Marcel J. C. Bijvelds

  2. Dalton Cardiovascular Research Center, Columbia, MO, USA

    Ashlee M. Strubberg, Jinghua Liu & Lane L. Clarke

  3. Department of Biomedical Sciences, University of Missouri, Columbia, MO, USA

    Ashlee M. Strubberg & Lane L. Clarke

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  1. Hugo R. de Jonge
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  2. Marcel J. C. Bijvelds
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  3. Ashlee M. Strubberg
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  4. Jinghua Liu
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  5. Lane L. Clarke
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Correspondence to Lane L. Clarke .

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  1. Department of Physiology, School of Biomedical Sciences, University of Otago, Dunedin, Otago, New Zealand

    Kirk L. Hamilton

  2. Department of Cell Biology, University of Pittsburgh, Pittsburgh, PA, USA

    Daniel C. Devor

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de Jonge, H.R., Bijvelds, M.J.C., Strubberg, A.M., Liu, J., Clarke, L.L. (2020). Organoids as a Model for Intestinal Ion Transport Physiology. In: Hamilton, K.L., Devor, D.C. (eds) Ion Transport Across Epithelial Tissues and Disease. Physiology in Health and Disease. Springer, Cham. https://doi.org/10.1007/978-3-030-55310-4_1

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