title: Expansion and differentiation of neural progenitors derived from the human adult enteric nervous system.
abstract: OBJECTIVE
  Neural stem and progenitor cells from the enteric nervous system have been proposed for use in cell-based therapies against specific neurogastrointestinal disorders. Recently, enteric neural progenitors were generated from human neonatal and early postnatal (until 5 years after birth) gastrointestinal tract tissues. We investigated the proliferation and differentiation of enteric nervous system progenitors isolated from human adult gastrointestinal tract.
  
  
  METHODS
  Human enteric spheroids were generated from adult small and large intestine tissues and then expanded and differentiated, depending on the applied cell culture conditions. For implantation studies, spheres were grafted into fetal slice cultures and embryonic aganglionic hindgut explants from mice. Differentiating enteric neural progenitors were characterized by 5-bromo-2-deoxyuridine labeling, in situ hybridization, immunocytochemistry, quantitative real-time polymerase chain reaction, and electrophysiological studies.
  
  
  RESULTS
  The yield of human neurosphere-like bodies was increased by culture in conditional medium derived from fetal mouse enteric progenitors. We were able to generate proliferating enterospheres from adult human small or large intestine tissues; these enterospheres could be subcultured and maintained for several weeks in vitro. Spheroid-derived cells could be differentiated into a variety of neuronal subtypes and glial cells with characteristics of the enteric nervous system. Experiments involving implantation into organotypic intestinal cultures showed the differentiation capacity of neural progenitors in a 3-dimensional environment.
  
  
  CONCLUSIONS
  It is feasible to isolate and expand enteric progenitor cells from human adult tissue. These findings offer new strategies for enteric stem cell research and future cell-based therapies.
authors: Translational Centre for Regenerative Medicine, University of Leipzig, Leipzig, Germany.
authors: Marco
authors: M
authors: Metzger
authors: Petra M
authors: PM
authors: Bareiss
authors: Timm
authors: T
authors: Danker
authors: Silvia
authors: S
authors: Wagner
authors: Joerg
authors: J
authors: Hennenlotter
authors: Elke
authors: E
authors: Guenther
authors: Florian
authors: F
authors: Obermayr
authors: Arnulf
authors: A
authors: Stenzl
authors: Alfred
authors: A
authors: Koenigsrainer
authors: Thomas
authors: T
authors: Skutella
authors: Lothar
authors: L
authors: Just
mesh_terms: D000328:Adult; D053687:Adult Stem Cells / Q000502:physiology*; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000378:metabolism; D004622:Embryo, Mammalian / Q000378:metabolism; D017615:Enteric Nervous System / Q000166:cytology / Q000502:physiology*; D005260:Female; D005333:Fetus / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007422:Intestines / Q000196:embryology / Q000294:innervation*; D008297:Male; D008564:Membrane Potentials; D051379:Mice; D008810:Mice, Inbred C57BL; D008875:Middle Aged; D009457:Neuroglia / Q000502:physiology*; D009474:Neurons / Q000502:physiology*; D018408:Patch-Clamp Techniques; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018874:Spheroids, Cellular; D013997:Time Factors; D046509:Tissue Culture Techniques
chemical_list: D017077:Culture Media, Conditioned; D012333:RNA, Messenger