Isolation of Enteric Nervous System Progenitor Cells from the Aganglionic Gut of Patients with Hirschsprung's Disease

Wilkinson, David J ORCID: 0000-0001-5165-8230, Bethell, George S, Shukla, Rajeev, Kenny, Simon E and Edgar, David H (2015) Isolation of Enteric Nervous System Progenitor Cells from the Aganglionic Gut of Patients with Hirschsprung's Disease. PLOS ONE, 10 (5). e0125724-.

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Abstract

Enteric nervous system progenitor cells isolated from postnatal human gut and cultured as neurospheres can then be transplanted into aganglionic gut to restore normal patterns of contractility. These progenitor cells may be of future use to treat patients with Hirschprung's disease, a congenital condition characterized by hindgut dysmotility due to the lack of enteric nervous system ganglia. Here we demonstrate that progenitor cells can also be isolated from aganglionic gut removed during corrective surgery for Hirschsprung's disease. Although the enteric nervous system marker calretinin is not expressed in the aganglionic gut region, de novo expression is initiated in cultured neurosphere cells isolated from aganglionic Hirschsprung bowel. Furthermore, expression of the neural markers NOS, VIP and GFAP also increased during culture of aganglionic gut neurospheres which we show can be transplantation into cultured embryonic mouse gut explants to restore a normal frequency of contractility. To determine the origin of the progenitor cells in aganglionic region, we used fluorescence-activated cell sorting to demonstrate that only p75-positive neural crest-derived cells present in the thickened nerve trunks characteristic of the aganglionic region of Hirschsprung gut gave rise to neurons in culture. The derivation of enteric nervous system progenitors in the aganglionic gut region of Hirschprung's patients not only means that this tissue is a potential source of cells for future autologous transplantation, but it also raises the possibility of inducing the differentiation of these endogenous cells in situ to compensate for the aganglionosis.

Item Type:	Article
Uncontrolled Keywords:	Intestine, Large, Enteric Nervous System, Animals, Humans, Mice, Hirschsprung Disease, Glial Fibrillary Acidic Protein, Vasoactive Intestinal Peptide, Flow Cytometry, Cell Culture Techniques, Cell Separation, Cell Differentiation, Nitric Oxide Synthase Type I, Adult Stem Cells, Neural Stem Cells, Heterografts, Biomarkers
Subjects:	?? QP ??
Depositing User:	Symplectic Admin
Date Deposited:	06 Aug 2015 16:12
Last Modified:	15 Dec 2022 09:55
DOI:	10.1371/journal.pone.0125724
Publisher's Statement :	Copyright: © 2015 Wilkinson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/2019182