title: Isolation of Enteric Nervous System Progenitor Cells from the Aganglionic Gut of Patients with Hirschsprung's Disease.
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.
authors.affiliation: University of Liverpool Institute of Translational Medicine, Liverpool, United Kingdom.
authors.forename: David J
authors.initials: DJ
authors.lastname: Wilkinson
authors.affiliation: University of Liverpool Institute of Translational Medicine, Liverpool, United Kingdom.
authors.forename: George S
authors.initials: GS
authors.lastname: Bethell
authors.affiliation: Department of Pathology, Alder Hey Children's Hospital NHS Foundation Trust, Liverpool, United Kingdom.
authors.forename: Rajeev
authors.initials: R
authors.lastname: Shukla
authors.affiliation: University of Liverpool Institute of Translational Medicine, Liverpool, United Kingdom; Department of Paediatric Surgery, Alder Hey Children's Hospital NHS Foundation Trust, Liverpool, United Kingdom.
authors.forename: Simon E
authors.initials: SE
authors.lastname: Kenny
authors.affiliation: University of Liverpool Institute of Translational Medicine, Liverpool, United Kingdom.
authors.forename: David H
authors.initials: DH
authors.lastname: Edgar
mesh_terms: D053687:Adult Stem Cells / Q000378:metabolism / Q000473:pathology* / Q000637:transplantation; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002469:Cell Separation; D017615:Enteric Nervous System / Q000378:metabolism / Q000473:pathology*; D005434:Flow Cytometry; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D064593:Heterografts; D006627:Hirschsprung Disease / Q000378:metabolism / Q000473:pathology* / Q000628:therapy; D006801:Humans; D007420:Intestine, Large / Q000294:innervation / Q000473:pathology; D051379:Mice; D058953:Neural Stem Cells / Q000378:metabolism / Q000473:pathology* / Q000637:transplantation; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism
chemical_list: D015415:Biomarkers; D005904:Glial Fibrillary Acidic Protein; D014660:Vasoactive Intestinal Peptide; C496320:NOS1 protein, human; D052248:Nitric Oxide Synthase Type I