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Neonatal intestinal organoids as an ex vivo approach to study early intestinal epithelial disorders

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Abstract

Background

Adult intestinal organoids have been used to study ex vivo intestinal injury in adulthood. However, the neonatal intestinal epithelium has many unique features that are different from adult mature intestine. Establishing a neonatal ex vivo organoid model is essential to study the epithelial physiology in early postnatal development and to investigate derangements associated with disease processes during the neonatal period like necrotizing enterocolitis (NEC).

Methods

Fresh and frozen terminal ileum was harvested from mice pups on postnatal day 9. Crypts were isolated and organoids were cultured. Organoids were exposed to hypoxia and lipopolysaccharide (LPS) for 48 h to induce epithelial injury. Inflammatory cytokines and tight junction proteins were evaluated.

Results

Robust intestinal organoids can be formed from both fresh and frozen intestinal tissue of neonatal mice pups. Hypoxia and LPS administration induced intestinal inflammation and disrupted tight junctions in these neonatal intestinal organoids.

Conclusions

We have established a novel method to grow organoids from neonatal intestine. We demonstrated that these organoids respond to the injury occurring during neonatal intestinal diseases such as NEC by increasing the organoid inflammation and by disrupting the organoid barrier function. Organoids provide an ex vivo platform to study intestinal physiology and pathology during the neonatal period.

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Acknowledgements

This work is supported by a Canadian Institutes of Health Research (CIHR) Foundation Grant 353857. AP is the Robert M. Filler Chair of Surgery, The Hospital for Sick Children (HSC). BL is the recipient of HSC Restracomp Fellowship.

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Authors and Affiliations

  1. Division of General and Thoracic Surgery, Translational Medicine Program, The Hospital for Sick Children, 555 University Avenue, Toronto, Ontario, M5G 1X8, Canada

    Bo Li, Carol Lee, Marissa Cadete, Hiromu Miyake, Dorothy Lee & Agostino Pierro

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  1. Bo Li
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  2. Carol Lee
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  3. Marissa Cadete
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  4. Hiromu Miyake
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  5. Dorothy Lee
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  6. Agostino Pierro
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Contributions

BL, CL, and AP designed experiments; CL, MC, HM, DL, and RW performed experiments; BL, CL, and MC wrote the manuscript; AP provided advice and supervision; all the authors reviewed and revised the manuscript.

Corresponding author

Correspondence to Agostino Pierro.

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The authors declare that they have no competing interest.

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Li, B., Lee, C., Cadete, M. et al. Neonatal intestinal organoids as an ex vivo approach to study early intestinal epithelial disorders. Pediatr Surg Int 35, 3–7 (2019). https://doi.org/10.1007/s00383-018-4369-3

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