Enteric nervous system stem cells derived from human gut mucosa for the treatment of aganglionic gut disorders.
OBJECTIVE
Enteric nervous system stem cells (ENSSCs) provide potential therapeutic tools to replenish absent ganglia in Hirschsprung's disease. Although full-thickness human postnatal gut tissue can be used to generate ENSSCs, reliance on its harvesting from surgical resection poses significant practical limitations. This study aimed to explore whether gut tissue obtained utilizing minimally invasive routine endoscopy techniques could be used to generate ENSSCs and whether such cells retain the potential to generate an ENS upon transplantation into aganglionic gut.


METHODS
Postnatal human gut mucosal tissue obtained from children undergoing gastrointestinal endoscopy was used to generate cell cultures in which ENSSCs were contained within neurosphere-like bodies (NLBs). These NLBs were characterized by immunostaining, and their potential to generate components of the ENS, in vitro and upon transplantation into models of aganglionic gut, was examined.


RESULTS
Gut mucosal biopsy specimens were obtained from 75 children (age, 9 months-17 years). The biopsy specimens contained neural cells and ENSSCs and, on culturing, generated characteristic NLBs at all ages examined. Postnatal mucosa-derived NLBs contained cells that, akin to their embryonic counterparts, were proliferating, expressed ENSSC markers, were bipotent, and capable of generating large colonies in clonogenic cultures and multiple ENS neuronal subtypes. Upon transplantation, cells from NLBs colonized cultured recipient aganglionic chick and human hindgut to generate ganglia-like structures and enteric neurons and glia.


CONCLUSIONS
The results represent a significant practical advance toward the development of definitive cell replenishment therapies for ENS disorders such as Hirschsprung's disease.
Metzger
Marco
M
Gastroenterology, Institute of Child Health, University College London, London, United Kingdom.
Caldwell
Claire
C
Barlow
Amanda J
AJ
Burns
Alan J
AJ
Thapar
Nikhil
N
D000293:Adolescent; D049109:Cell Proliferation; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D017615:Enteric Nervous System / Q000166:cytology* / Q000196:embryology; D005260:Female; D041981:Gastrointestinal Tract / Q000166:cytology / Q000502:physiology; D006627:Hirschsprung Disease / Q000473:pathology / Q000628:therapy*; D006801:Humans; D007223:Infant; D007413:Intestinal Mucosa / Q000166:cytology / Q000637:transplantation*; D008297:Male; D008856:Microscopy, Fluorescence; D009092:Mucous Membrane / Q000637:transplantation; D012038:Regeneration; D012494:Sampling Studies; D012680:Sensitivity and Specificity; D033581:Stem Cell Transplantation / Q000379:methods*