title: Skin-derived precursors generate enteric-type neurons in aganglionic jejunum.
abstract: OBJECTIVE
  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 65-85days 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Surgery, Division of Pediatric Surgery, University of California, Los Angeles, Los Angeles, CA 90095-1749, USA.
authors.forename: Justin P
authors.initials: JP
authors.lastname: Wagner
authors.affiliation: Department of Surgery, Division of Pediatric Surgery, University of California, Los Angeles, Los Angeles, CA 90095-1749, USA.
authors.forename: Veronica F
authors.initials: VF
authors.lastname: Sullins
authors.affiliation: Department of Surgery, Division of Pediatric Surgery, University of California, Los Angeles, Los Angeles, CA 90095-1749, USA; Department of Bioengineering, University of California, Los Angeles, Los Angeles, CA 90095-7098, USA. Electronic address: jdunn@mednet.ucla.edu.
authors.forename: James C Y
authors.initials: JC
authors.lastname: Dunn
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation; D002478:Cells, Cultured; D004195:Disease Models, Animal; D017615:Enteric Nervous System / Q000166:cytology*; D005260:Female; D005726:Ganglia, Parasympathetic / Q000166:cytology*; D006627:Hirschsprung Disease / Q000473:pathology / Q000628:therapy*; D007583:Jejunum / Q000294:innervation / Q000473:pathology*; D009432:Neural Crest / Q000166:cytology*; D051381:Rats; D011917:Rats, Inbred Lew; D012867:Skin / Q000166:cytology*; D033581:Stem Cell Transplantation / Q000379:methods*
keywords: Aganglionosis; Cell-based therapy; Hirschsprung’s disease; Regenerative medicine; Stem cell