Determining Intestinal Permeability Using Lucifer Yellow in an Apical-Out Enteroid Model

作者： Heather Liebe1, Camille Schlegel2, Xue Cai2, Alena Golubkova1, Tyler Leiva1, William L. Berry3, Catherine J. Hunter1 1Division of Pediatric Surgery,Oklahoma Children's Hospital, 2Oklahoma University Health Sciences Center, 3Department of Surgery,University of Oklahoma Health Sciences Center

简介：

Overview

This protocol describes a method using lucifer yellow in an apical-out enteroid model to assess intestinal permeability. It is particularly relevant for studying inflammatory bowel diseases like necrotizing enterocolitis.

Key Study Components

Area of Science

• Neuroscience
• Biology
• Gastroenterology

Background

• Lucifer yellow is a fluorescent dye used to evaluate permeability.
• Enteroids are 3D models that mimic intestinal tissue.
• Paracellular permeability is crucial for understanding intestinal health.
• This method can be applied to various intestinal diseases.

Purpose of Study

• To develop a reliable method for assessing intestinal permeability.
• To utilize enteroid models for studying human intestinal diseases.
• To investigate the mechanisms underlying necrotizing enterocolitis.

Methods Used

• Application of lucifer yellow to the media of apical-out enteroids.
• Growing enteroids embedded in BMM for 7-10 days.
• Using 50% LWRN conditioned media for enteroid generation.
• Conducting quantitative and qualitative analysis of permeability pathways.

Main Results

• Demonstrated a straightforward technique for permeability assessment.
• Showed potential applications in various intestinal diseases.
• Highlighted the role of enteroids in studying disease mechanisms.
• Involved collaboration among research scholars and assistants.

Conclusions

• The lucifer yellow method is effective for studying intestinal permeability.
• Apical-out enteroids provide a valuable model for research.
• This technique can enhance understanding of intestinal diseases.

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