title: Isogenic enteric neural progenitor cells can replace missing neurons and glia in mice with Hirschsprung disease.
abstract: BACKGROUND
  Transplanting autologous patient-derived enteric neuronal stem/progenitor cells (ENSCs) is an innovative approach to replacing missing enteric neurons in patients with Hirschsprung disease (HSCR). Using autologous cells eliminates immunologic and ethical concerns raised by other cell sources. However, whether postnatal aganglionic bowel is permissive for transplanted ENSCs and whether ENSCs from HSCR patients can be successfully isolated, cultured, and transplanted in vivo remains unknown.
  
  
  METHODS
  ENSCs isolated from the ganglionic intestine of Ednrb(-/-) mice (HSCR-ENSCs) were characterized immunohistochemically and evaluated for their capacity to proliferate and differentiate in vitro. Fluorescently labeled ENSCs were co-cultured ex vivo with aganglionic Ednrb(-/-) colon. For in vivo transplantation, HSCR-ENSCs were labeled with lentivirus expressing green fluorescent protein (GFP) and implanted into aganglionic embryonic chick gut in ovo and postnatal aganglionic Ednrb(-/-) rectum in vivo.
  
  
  RESULTS
  HSCR-ENSCs maintain normal capacity self-renewal and neuronal differentiation. Moreover, the Ednrb(-/-) aganglionic environment is permissive to engraftment by wild-type ENSCs ex vivo and supports migratrion and neuroglial differentiation of these cells following transplantation in vivo. Lentiviral GFP-labeled HSCR-ENSCs populated embryonic chick hindgut and postnatal colon of Ednrb(-/-) HSCR, with cells populating the intermuscular layer and forming enteric neurons and glia.
  
  
  CONCLUSIONS
  ENSCs can be isolated and cultured from mice with HSCR, and transplanted into the aganglionic bowel of HSCR littermates to generate enteric neuronal networks. These results in an isogenic model establish the potential of using autologous-derived stem cells to treat HSCR and other intestinal neuropathies.
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
authors.forename: R
authors.initials: R
authors.lastname: Hotta
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.|Department of Surgery, University of California San Francisco, San Francisco, CA, USA.
authors.forename: L S
authors.initials: LS
authors.lastname: Cheng
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.
authors.forename: H K
authors.initials: HK
authors.lastname: Graham
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.|Department of Pediatric Surgery, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China.
authors.forename: W
authors.initials: W
authors.lastname: Pan
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.|Department of Human Morphology and Developmental Biology, Faculty of Medicine, Semmelweis University, Budapest, Hungary.
authors.forename: N
authors.initials: N
authors.lastname: Nagy
authors.affiliation: Department of Pediatric Gastroenterology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.|Pediatric Neurogastroenterology Program, Massachusetts General Hospital, MA, USA.
authors.forename: J
authors.initials: J
authors.lastname: Belkind-Gerson
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA, USA.|Pediatric Neurogastroenterology Program, Massachusetts General Hospital, MA, USA.
authors.forename: A M
authors.initials: AM
authors.lastname: Goldstein
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D006627:Hirschsprung Disease; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D058953:Neural Stem Cells / Q000637:transplantation*; D009457:Neuroglia / Q000166:cytology*; D009474:Neurons / Q000166:cytology*; D033581:Stem Cell Transplantation / Q000379:methods*; D014185:Transplantation, Isogeneic / Q000379:methods
keywords: Hirschsprung disease; Lentivirus; cell therapy; endothelin receptor B; enteric nervous system