Expansion and differentiation of neural progenitors derived from the human adult enteric nervous system.
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.
Metzger
Marco
M
Translational Centre for Regenerative Medicine, University of Leipzig, Leipzig, Germany.
Bareiss
Petra M
PM
Danker
Timm
T
Wagner
Silvia
S
Hennenlotter
Joerg
J
Guenther
Elke
E
Obermayr
Florian
F
Stenzl
Arnulf
A
Koenigsrainer
Alfred
A
Skutella
Thomas
T
Just
Lothar
L
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
D017077:Culture Media, Conditioned; D012333:RNA, Messenger