Isolation of high-purity myenteric plexus from adult human and mouse gastrointestinal tract

Scientific Reports
David GrundmannKarl-Herbert Schäfer

Abstract

The enteric nervous system (ENS) orchestrates a broad range of important gastrointestinal functions such as intestinal motility and gastric secretion. The ENS can be affected by environmental factors, diet and disease. Changes due to these alterations are often hard to evaluate in detail when whole gut samples are used. Analyses based on pure ENS tissue can more effectively reflect the ongoing changes during pathological processes. Here, we present an optimized approach for the isolation of pure myenteric plexus (MP) from adult mouse and human. To do so, muscle tissue was individually digested with a purified collagenase. After incubation and a gentle mechanical disruption step, MP networks could be collected with anatomical integrity. These tissues could be stored and used either for immediate genomic, proteomic or in vitro approaches, and enteric neurospheres could be generated and differentiated. In a pilot experiment, the influence of bacterial lipopolysaccharide on human MP was analyzed using 2-dimensional gel electrophoresis. The method also allows investigation of factors that are secreted by myenteric tissue in vitro. The isolation of pure MP in large amounts allows new analytical approaches that can provide a new persp...Continue Reading

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Citations

Dec 27, 2015·Journal of Neuroscience Methods·George WahbaSarah Schock
Jan 1, 2016·Stem Cell Research & Therapy·Rhian StavelyKulmira Nurgali
Apr 10, 2016·Developmental Biology·Alan J BurnsPieter Vanden Berghe
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Apr 12, 2019·Journal of Virology·Andrew K HastingsErol Fikrig
Aug 22, 2017·Frontiers in Physiology·Dominik SchreiberKarl-Herbert Schäfer
Jun 19, 2020·Npj Regenerative Medicine·Suradip DasD Kacy Cullen
Nov 13, 2015·Current Opinion in Clinical Nutrition and Metabolic Care·Stijn SoenenMichael Horowitz
Nov 30, 2020·Journal of Neuroscience Methods·Zitong HuangBo Tan
Jan 31, 2021·Nature Reviews. Gastroenterology & Hepatology·Beate NieslerKarl-Herbert Schäfer
Feb 14, 2021·Npj Regenerative Medicine·Suradip DasD Kacy Cullen
Jun 4, 2021·Molecular Neurodegeneration·Manuela GriesKarl-Herbert Schäfer
Jul 9, 2021·Neurogastroenterology and Motility : the Official Journal of the European Gastrointestinal Motility Society·Simone L SchonkerenVeerle Melotte

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Methods Mentioned

BETA
transgenic
scanning electron microscopy
PCR
ELISAs
biopsies
chip
FCS
ELISA

Software Mentioned

Zeiss
Liberase
CellObserver
TCX
MEA Setup
- Rack
- MEA
MEA
Change
Control

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