title: Optimizing neurogenic potential of enteric neurospheres for treatment of neurointestinal diseases.
abstract: Enteric neurospheres derived from postnatal intestine represent a promising avenue for cell replacement therapy to treat Hirschsprung disease and other neurointestinal diseases. We describe a simple method to improve the neuronal yield of spontaneously formed gut-derived neurospheres.
  
  
  
  Enteric neurospheres were formed from the small and large intestines of mouse and human subjects. Neurosphere size, neural crest cell content, cell migration, neuronal differentiation, and neuronal proliferation in culture were analyzed. The effect of supplemental neurotrophic factors, including glial cell line-derived neurotrophic factor (GDNF) and endothelin-3, was also assessed.
  
  
  
  Mouse small intestine-derived neurospheres contained significantly more P75-expressing neural crest-derived cells (49.9 ± 15.3% versus 21.6 ± 11.9%, P < 0.05) and gave rise to significantly more Tuj1-expressing neurons than colon-derived neurospheres (69.9 ± 8.6% versus 46.2 ± 15.6%, P < 0.05). A similar pattern was seen in neurospheres isolated from human small and large intestine (32.6 ± 17.5% versus 10.2 ± 8.2% neural crest cells, P < 0.05; 29.7 ± 16.4% versus 16.0 ± 13.5% enteric neurons, P < 0.05). The addition of GDNF to the culture media further improved the neurogenic potential of small intestinal neurospheres (75.9 ± 4.0% versus 67.8 ± 5.8%, P < 0.05) whereas endothelin-3 had no effect.
  
  
  
  Enteric neurospheres formed from small intestine and supplemented with GDNF yield an enriched population of neural crest-derived progenitor cells and give rise to a high density of enteric neurons.
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, 55 Fruit St., Boston, MA, 02114, USA.|Department of Surgery, University of California San Francisco, 500 Parnassus Ave., San Francisco, CA, 94143, USA.
authors.forename: Lily S
authors.initials: LS
authors.lastname: Cheng
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, 55 Fruit St., Boston, MA, 02114, USA.
authors.forename: Hannah K
authors.initials: HK
authors.lastname: Graham
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, 55 Fruit St., Boston, MA, 02114, USA.|Department of Pediatric Surgery, Xinhua Hospital, Shanghai Jiaotong University School of Medicine, 280 Chongqing S Rd, Huangpu, Shanghai, China.
authors.forename: Wei Hua
authors.initials: WH
authors.lastname: Pan
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, 55 Fruit St., Boston, MA, 02114, USA.|Department of Anatomy, Histology and Embryology, Faculty of Medicine, Semmelweis University, Tuzolto St. 58, Budapest 1094, Hungary.
authors.forename: Nandor
authors.initials: N
authors.lastname: Nagy
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, 55 Fruit St., Boston, MA, 02114, USA.|Laboratorio de Genética y Biomarcadores, Instituto Nacional de Salud Pública, Av Universidad 655, Santa María Ahuacatitlán, 62100 Cuernavaca, Mor., Mexico.
authors.forename: Alfonso
authors.initials: A
authors.lastname: Carreon-Rodriguez
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, 55 Fruit St., Boston, MA, 02114, USA.
authors.forename: Allan M
authors.initials: AM
authors.lastname: Goldstein
authors.affiliation: Department of Pediatric Surgery, Massachusetts General Hospital, Harvard Medical School, 55 Fruit St., Boston, MA, 02114, USA.
authors.forename: Ryo
authors.initials: R
authors.lastname: Hotta
mesh_terms: D000293:Adolescent; D000818:Animals; D002454:Cell Differentiation; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D002648:Child; D017615:Enteric Nervous System / Q000166:cytology* / Q000502:physiology; D005260:Female; D005767:Gastrointestinal Diseases / Q000628:therapy; D006627:Hirschsprung Disease / Q000628:therapy; D006801:Humans; D007223:Infant; D007420:Intestine, Large / Q000166:cytology / Q000502:physiology; D007421:Intestine, Small / Q000166:cytology / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D058953:Neural Stem Cells / Q000502:physiology / Q000637:transplantation*; D055495:Neurogenesis / Q000502:physiology*; D055815:Young Adult
keywords: Endothelin-3; Enteric nervous system; Enteric neural stem cells; Glial cell line-derived neurotrophic factor; Neurospheres