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Related Experiment Video

Updated: Aug 7, 2025

In Vitro Apical-Out Enteroid Model of Necrotizing Enterocolitis
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In Vitro Apical-Out Enteroid Model of Necrotizing Enterocolitis

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Apical-Out Enteroids as an Innovative Model for Necrotizing Enterocolitis.

Heather Liebe1, Camille Schlegel2, Xue Cai2

  • 1Division of Pediatric Surgery, Oklahoma Children's Hospital, Oklahoma City, Oklahoma.

The Journal of Surgical Research
|March 14, 2023
PubMed
Summary

Apical-out enteroids offer a novel model for necrotizing enterocolitis (NEC) in premature infants. This model demonstrates increased inflammation and gut barrier changes, enabling better study of pathogen interactions in NEC.

Keywords:
Apical-outEnteroidsModelNecrotizing enterocolitis

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Area of Science:

  • Gastroenterology
  • Neonatal Research
  • Cell Biology

Background:

  • Necrotizing enterocolitis (NEC) is a critical gastrointestinal disease affecting premature neonates.
  • Existing human infant intestinal enteroid models lack direct apical access, limiting physiological study.
  • Investigating NEC pathogenesis requires models that mimic the intestinal epithelial barrier's interaction with luminal factors.

Purpose of the Study:

  • To develop and validate an apical-out (AO) enteroid model for studying necrotizing enterocolitis (NEC).
  • To assess inflammatory responses and gut barrier integrity in AO enteroids under NEC-relevant conditions.
  • To enable a more biologically relevant investigation of pathogen-epithelial interactions in NEC.

Main Methods:

  • Human neonatal intestinal tissue was used to generate enteroids.
  • Enteroids were cultured and everted to achieve an apical-out (AO) configuration.
  • AO enteroids were treated with lipopolysaccharide and hypoxia to simulate NEC conditions, followed by analysis of inflammatory markers and tight junction proteins.

Main Results:

  • Apical-out configuration was confirmed via immunofluorescence.
  • Treated AO enteroids showed significantly increased TNF-α and TLR4 expression.
  • Significant downregulation of tight junction proteins (Occludin, Claudins, ZO-1) was observed in treated AO enteroids, indicating barrier disruption.

Conclusions:

  • Apical-out enteroids provide an innovative and relevant model for NEC research.
  • This model exhibits increased inflammation and gut barrier restructuring characteristic of NEC.
  • AO enteroids facilitate biologically relevant studies of pathogen-intestinal barrier interactions in NEC.