Characterisation and transplantation of enteric nervous system progenitor cells

Abstract

Aims: Enteric nervous system (ENS) progenitor cells have been postulated to be an appropriate source of cells for the treatment of Hirschsprung's disease. In order for this to be successful, the techniques previously used for the isolation of rodent ENS progenitor cells need to be adapted for postnatal human tissue. In this paper, we describe a method suitable for the preparation of both mouse and human postnatal ENS progenitor cells and assess their transplantation potential. Method: Single cell suspensions were isolated from 11.5 days post-coitum embryonic mouse caecum and postnatal human myenteric plexus. These cells were cultured under non-adherent conditions to generate neurospheres which were implanted into aganglionic embryonic mouse hindgut explants. Cell proliferation, migration and differentiation were observed using immunofluorescence microscopy. Results: Neurospheres generated from both mouse and human tissues contained proliferating neural crestderived cells that could be expanded in tissue culture to generate both glial cells and neurons. When implanted into aganglionic murine gut, cells migrated from the neurospheres using pathways appropriate for cells derived from the neural crest, and differentiated to become glia and neurons expressing neuronal phenotypic markers characteristic of the ENS including nitric oxide synthase and vasoactive intestinal polypeptide. Conclusion: We have developed a technique for the isolation and expansion of ENS progenitor cells from human neonates. These cells have the ability to differentiate into neurons and glia when transplanted into aganglionic gut, this demonstration being a necessary first step for their autologous transplantation in the treatment of Hirschsprung's disease.

Involvement of DNMT3B in the pathogenesis of Hirschsprung disease and its possible role as a regulator of neurogenesis in the human enteric nervous system

“…These NLBs have been used as a source of enteric precursor cells that can be transplanted to restore the contractile properties of aganglionic bowel by neurally mediated mechanisms. [25][26][27][28] In the present study, we have used such NLBs to search for new genes that could be implicated in ENS formation and in the pathogenesis of HSCR. Given that neural crest pluripotency is essential for proper ENS development in particular, we have studied the expression profiles of pluripotency-associated genes in enteric neural precursors.…”

Isolation and Characterization of Neural Crest Stem Cells Derived From In Vitro–Differentiated Human Embryonic Stem Cells

“…Thus, the feasibility of acquiring primary human neural crest cells from embryonic tissue is severely challenged by both ethical and technical issues-in fact, neural crest induction and migration is underway long before most women even realize that they are pregnant. Although NCSCs have been isolated from some human adult tissues, they are exceedingly rare and do not possess the same selfrenewal and pluripotency of early embryonic NCSCs [1,11,12]. Therefore, to fully advance in vitro studies of human neural crest development alternate renewable sources of precursor cells will be required.…”

Tissue Engineering Functional Gastrointestinal Regions: The Importance of Stem and Progenitor Cells

“…The ENS, a critical component of functional intestine, has reproducibly been expanded from ENS progenitors in vitro (Almond et al 2007;Bitar and Raghavan 2014). Therefore, ENS progenitors may be propagated as isolated intestinal components, or together with muscle and epithelium for the formation of functional bowel.…”