Nestin‐expressing cells in the gut give rise to enteric neurons and glial cells

Abstract

Background Neuronal stem cells (NSCs) are promising for neurointestinal disease therapy. While NSCs have been isolated from intestinal musclularis, their presence in mucosa has not been well described. Mucosa-derived NSCs are accessible endoscopically and could be used autologously. Brain-derived Nestin-positive NSCs are important in endogenous repair and plasticity. The aim was to isolate and characterize mucosa-derived NSCs, determine their relationship to Nestin-expressing cells and demonstrate capacity to produce neuroglial networks in vitro and in vivo. Methods Neurospheres were generated from periventricular brain, colonic muscularis (Musc), and mucosa-submucosa (MSM) of mice expressing green fluorescent protein (GFP) controlled by the Nestin promoter (Nestin-GFP). NSCs were also grown as adherent colonies from intestinal mucosal organoids. Their differentiation potential was assessed by immunohistochemistry using glial and neuronal markers. Brain and gut derived neurospheres were transplanted into explants of chick embryonic aneural hindgut to determine their fate. Results Musc- and MSM-derived neurospheres expressed Nestin and gave rise to cells of neuronal, glial and mesenchymal lineage. While Nestin expression in tissue was mostly limited to glia colabelled with glial fibrillary acid protein (GFAP), neurosphere-derived neurons and glia both expressed Nestin in vitro, suggesting Nestin+/GFAP+ glial cells may give rise to new neurons. Moreover, following transplantation into aneural colon, brain- and gut-derived NSCs were able to differentiate into neurons. Conclusions Nestin-expressing intestinal NSCs cells give rise to neurospheres, differentiate into neuronal, glial and mesenchymal lineages in vitro, generate neurons in vivo and can be isolated from mucosa. Further studies are needed exploring their potential for treating neuropathies.

Delivery of enteric neural progenitors with 5-HT4 agonist-loaded nanoparticles and thermosensitive hydrogel enhances cell proliferation and differentiation following transplantation in vivo

“…Immunohistochemistry was performed as previously described [40]. Cells or tissues were fixed, washed, and permeabilized with 0.1% Triton X-100 for 30 minutes.…”

Migration and differentiation of transplanted enteric neural crest-derived cells in murine model of Hirschsprung’s disease

“…Characterization of isolated ENS stem/progenitor cells from P0-Cre/EGFP mouse gut in the current study shows they express markers of neural crest (p75 and Sox10) and neural stem/progenitor cells (Nestin and Musashi1). It has been reported that Nestin positive cells in enteric ganglia of mouse and human (Azan et al 2011) form NLBs and give rise to neurons and glia in culture (Suarez-Rodriguez and BelkindGerson 2004;Belkind-Gerson et al 2013). Musashi1, a well-known marker for stem cells in nervous system (Sakakibara et al 1996;Okano et al 2002) and intestine (Potten et al 2003), plays a key role in maintaining undifferentiated status of neural stem cells (Okano et al 2002).…”

“…These cells can give rise to neurons and glial cells when grafted into explants of chick, mouse or human gut (Mosher et al 2007;Metzger et al 2009;Lindley et al 2008;Belkind-Gerson et al 2013) or postnatal mouse gut in vivo (Hotta et al 2013). Moreover, a recent human study has shown that enteric neuronal stem/progenitor cells can be obtained from mucosal biopsy specimens taken during routine endoscopy from children with Hirschsprung's disease (Metzger et al 2009), demonstrating the potential for the use of patient-derived cells in the future for autologous cell replacement that circumvent ethical and immunological challenges.…”

601 The Appendix As a Viable Source of Neural Progenitor Cells to Functionally Innervate Bioengineered Gastrointestinal Smooth Muscle Tissues

“…Well-established protocols have been developed to isolate enteric NPCs from the GI tract [21,22]. When transplanted into rodents, these cells differentiated into neurons that reestablished motility or function [23,24].…”