PMID- 17460755
OWN - NLM
STAT- MEDLINE
DCOM- 20100419
LR  - 20220410
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 2
IP  - 4
DP  - 2007 Apr 25
TI  - Enriched monolayer precursor cell cultures from micro-dissected adult mouse 
      dentate gyrus yield functional granule cell-like neurons.
PG  - e388
LID - e388
AB  - BACKGROUND: Stem cell cultures are key tools of basic and applied research in 
      Regenerative Medicine. In the adult mammalian brain, lifelong neurogenesis 
      originating from local precursor cells occurs in the neurogenic regions of the 
      hippocampal dentate gyrus. Despite widespread interest in adult hippocampal 
      neurogenesis and the use of mouse models to study it, no protocol existed for 
      adult murine long-term precursor cell cultures with hippocampus-specific 
      differentiation potential. METHODOLOGY/PRINCIPAL FINDINGS: We describe a new 
      strategy to obtain serum-free monolayer cultures of neural precursor cells from 
      microdissected dentate gyrus of adult mice. Neurons generated from these adherent 
      hippocampal precursor cell cultures expressed the characteristic markers like 
      transcription factor Prox1 and showed the TTX-sensitive sodium currents of mature 
      granule cells in vivo. Similar to granule cells in vivo, treatment with kainic 
      acid or brain derived neurotrophic factor (BDNF) elicited the expression of 
      GABAergic markers, further supporting the correspondence between the in vitro and 
      in vivo phenotype. When plated as single cells (in individual wells) or at lowest 
      density for two to three consecutive generations, a subset of the cells showed 
      self-renewal and gave rise to cells with properties of neurons, astrocytes and 
      oligodendrocytes. The precursor cell fate was sensitive to culture conditions 
      with their phenotype highly influenced by factors within the media (sonic 
      hedgehog, BMP, LIF) and externally applied growth factors (EGF, FGF2, BDNF, and 
      NT3). CONCLUSIONS/SIGNIFICANCE: We report the conditions required to generate 
      adult murine dentate gyrus precursor cell cultures and to analyze functional 
      properties of precursor cells and their differentiated granule cell-like progeny 
      in vitro.
FAU - Babu, Harish
AU  - Babu H
AD  - Max Delbruck Center for Molecular Medicine (MDC) Berlin-Buch, Berlin, Germany.
FAU - Cheung, Giselle
AU  - Cheung G
FAU - Kettenmann, Helmut
AU  - Kettenmann H
FAU - Palmer, Theo D
AU  - Palmer TD
FAU - Kempermann, Gerd
AU  - Kempermann G
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20070425
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
SB  - IM
MH  - Animals
MH  - Cell Differentiation
MH  - Cell Proliferation
MH  - Cells, Cultured
MH  - Dentate Gyrus/*cytology
MH  - Humans
MH  - Mice
MH  - Neurons/*cytology
MH  - Rats
PMC - PMC1849968
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2007/04/27 09:00
MHDA- 2007/04/27 09:01
PMCR- 2007/04/25
CRDT- 2007/04/27 09:00
PHST- 2006/12/19 00:00 [received]
PHST- 2007/03/27 00:00 [accepted]
PHST- 2007/04/27 09:00 [pubmed]
PHST- 2007/04/27 09:01 [medline]
PHST- 2007/04/27 09:00 [entrez]
PHST- 2007/04/25 00:00 [pmc-release]
AID - 06-PONE-RA-00505R1 [pii]
AID - 10.1371/journal.pone.0000388 [doi]
PST - epublish
SO  - PLoS One. 2007 Apr 25;2(4):e388. doi: 10.1371/journal.pone.0000388.