US EPA

2840254 

Journal Article 

Skin-derived precursors generate enteric-type neurons in aganglionic jejunum 

Wagner, JP; Sullins, VF; Dunn, JCY 

2014 

Yes 

Journal of Pediatric Surgery
ISSN: 0022-3468
EISSN: 1531-5037 

49 

12 

1809-1814 

25487489 

10.1016/j.jpedsurg.2014.09.023 

WOS:000345965000026 

Purpose: Skin-derived precursor cells (SKPs) may regenerate the enteric nervous system in Hirschsprung's disease. SKPs migrate and differentiate into myenteric ganglia in aganglionic intestine. We sought to characterize the time-course of SKP gangliogenesis and enteric neurotransmitter synthesis in vivo.



Methods: Adult Lewis rat jejunal segments were isolated and denervated with benzalkonium chloride (BAC). Denervation was evaluated by immunohistochemical (IHC) stains for markers of mature neuronal and glial cells. Green fluorescent protein (GFP)-expressing neonatal rat SKPs were cultured in neuroglial-selective medium. SKPs were transplanted into aganglionic segments 65-85 days after BAC treatment. IHC was performed to identify glia, neurons, and neurotransmitter synthesis in GFP + cells between post-transplant days 1 and 28.



Results: Aganglionosis was confirmed by IHC. On post-transplant days 1 and 2, GFP + cells were detected near injection sites within the muscularis propria. GFP + cell clusters were evident only between longitudinal and circular smooth muscle layers at post-transplant days 14, 21, and 28. These structures co-expressed markers of mature neurons and gliocytes. Several markers of neurotransmitter synthesis were detected in GFP + clusters at days 21 and 28.



Conclusion: SKPs are capable of enteric neuroglial differentiation in vivo. SKPs migrate to the intermuscular layer of aganglionic intestine within days of transplantation. Our observations suggest that SKPs are capable of generating enteric ganglia in aganglionic intestine. (C) 2014 Elsevier Inc. All rights reserved. 

Hirschsprung's disease; Aganglionosis; Regenerative medicine; Stem cell; Cell-based therapy