PMID- 16527901
OWN - NLM
STAT- MEDLINE
DCOM- 20060825
LR  - 20141120
IS  - 1066-5099 (Print)
IS  - 1066-5099 (Linking)
VI  - 24
IP  - 6
DP  - 2006 Jun
TI  - In vitro formation of enteric neural network structure in a gut-like organ 
      differentiated from mouse embryonic stem cells.
PG  - 1414-22
AB  - Using an embryoid body (EB) culture system, we developed a functional organ-like 
      cluster--a "gut"--from mouse embryonic stem (ES) cells (ES gut). Each ES gut 
      exhibited spontaneous contractions but did not exhibit distinct peristalsis-like 
      movements. In these spontaneously contracting ES guts, dense distributions of 
      interstitial cells of Cajal (c-kit [a transmembrane receptor that has tyrosine 
      kinase activity]-positive cells; gut pacemaker cells) and smooth muscle cells 
      were discernibly identified; however, enteric neural ganglia were absent in the 
      spontaneously differentiated ES gut. By adding brain-derived neurotrophic factor 
      (BDNF) only during EB formation, we for the first time succeeded in in vitro 
      formation of enteric neural ganglia with connecting nerve fiber tracts (enteric 
      nervous system [ENS]) in the ES gut. The ES gut with ENS exhibited strong 
      peristalsis-like movements. During EB culture in BDNF(+) medium, we detected each 
      immunoreactivity associated with the trk proto-oncogenes (trkB; BDNF receptors) 
      and neural crest marker, proto-oncogene tyrosine-protein kinase receptor ret 
      precursor (c-ret), p75, or sox9. These results indicated that the present ENS is 
      differentiated from enteric neural crest-derived cells. Moreover, focal 
      stimulation of ES guts with ENS elicited propagated increases in intracellular 
      Ca(2+) concentration ([Ca(2+)](i)) at single or multiple sites that were 
      attenuated by atropine or abolished by tetrodotoxin. These results suggest in 
      vitro formation of physiologically functioning enteric cholinergic excitatory 
      neurons. We for the first time succeeded in the differentiation of functional 
      neurons in ENS by exogenously adding BDNF in the ES gut, resulting in generation 
      of distinct peristalsis-like movements.
FAU - Takaki, Miyako
AU  - Takaki M
AD  - Department of Physiology II, Nara Medical University, School of Medicine, 
      Kashihara, Japan. mtakaki@naramed-u.ac.jp
FAU - Nakayama, Shinsuke
AU  - Nakayama S
FAU - Misawa, Hiromi
AU  - Misawa H
FAU - Nakagawa, Tadashi
AU  - Nakagawa T
FAU - Kuniyasu, Hiroki
AU  - Kuniyasu H
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20060309
PL  - England
TA  - Stem Cells
JT  - Stem cells (Dayton, Ohio)
JID - 9304532
RN  - 0 (Brain-Derived Neurotrophic Factor)
RN  - EC 2.7.10.1 (Proto-Oncogene Proteins c-ret)
RN  - EC 2.7.10.1 (Receptor, trkB)
SB  - IM
MH  - Animals
MH  - Brain-Derived Neurotrophic Factor/pharmacology
MH  - Calcium Signaling
MH  - Cell Differentiation/drug effects
MH  - Cells, Cultured
MH  - Embryo, Mammalian/cytology
MH  - Enteric Nervous System/cytology/drug effects/*embryology/metabolism
MH  - Gastrointestinal Motility
MH  - In Vitro Techniques
MH  - Mice
MH  - Nerve Net/cytology/drug effects/embryology/metabolism
MH  - Proto-Oncogene Proteins c-ret/metabolism
MH  - Receptor, trkB/metabolism
MH  - Stem Cells/*cytology/drug effects
EDAT- 2006/03/11 09:00
MHDA- 2006/08/26 09:00
CRDT- 2006/03/11 09:00
PHST- 2006/03/11 09:00 [pubmed]
PHST- 2006/08/26 09:00 [medline]
PHST- 2006/03/11 09:00 [entrez]
AID - 2005-0394 [pii]
AID - 10.1634/stemcells.2005-0394 [doi]
PST - ppublish
SO  - Stem Cells. 2006 Jun;24(6):1414-22. doi: 10.1634/stemcells.2005-0394. Epub 2006 
      Mar 9.