title: Transplanted skin-derived precursor stem cells generate enteric ganglion-like structures in vivo.
abstract: BACKGROUND
  Hirschsprung's disease is characterized by a developmental arrest of neural crest cell migration, causing distal aganglionosis. Transplanted cells derived from the neural crest may regenerate enteric ganglia in this condition. We investigated the potential of skin-derived precursor cells (SKPs) to engraft and to differentiate into enteric ganglia in aganglionic rat intestine in vivo.
  
  
  METHODS
  Adult Lewis rat jejunal segments were separated from intestinal continuity and treated with benzalkonium chloride to induce aganglionosis. Ganglia were identified via immunohistochemical stains for S100 and β-III tubulin (TUJ1). SKPs were procured from neonatal Lewis rats expressing enhanced green fluorescent protein (GFP) and cultured in neuroglial-selective media. SKP cell line expansion was quantified, and immunophenotypes were assessed by immunocytochemistry. Aganglionic segments underwent SKP transplantation 21-79days after benzalkonium chloride treatment. The presence of GFP+cells, mature neurons, and mature glia was evaluated at posttransplant days 1, 6, and 9.
  
  
  RESULTS
  Benzalkonium chloride-induced aganglionosis persisted for at least 85days. Prior to differentiation, SKPs expressed S100, denoting neural crest lineage, and nestin, a marker of neuronal precursors. Differentiated SKPs in vitro expressed GFAP, a marker of glial differentiation, as well as TUJ1 and several enteric neurotransmitters. After transplantation, GFP+structures resembling ganglia were identified between longitudinal and circular smooth muscle layers.
  
  
  CONCLUSIONS
  SKPs are capable of engraftment, migration, and differentiation within aganglionic rodent intestine in vivo. Differentiated SKPs generate structures that resemble enteric ganglia. Our observations suggest that SKPs represent a potential gangliogenic therapeutic agent for Hirschsprung's disease.
authors.affiliation: Department of Surgery, Division of Pediatric Surgery, University of California, Los Angeles, Los Angeles, CA 90095-1749, USA. Electronic address: jwagner@mednet.ucla.edu.
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; D005726:Ganglia, Parasympathetic / Q000166:cytology*; D006627:Hirschsprung Disease / Q000473:pathology / Q000628:therapy*; D008297:Male; 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; Neural crest; Regenerative medicine; Skin-derived precursor cells; Stem cell
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