title: [Organotypic slice culture of neonatal rat cortex and induced neural stem cell differentiation].
abstract: OBJECTIVE
  To establish a method for organotypic slice culture of neonatal rat cortex in a modified condition and investigate the effect of spatial signals on neural stem cell (NSC) differentiation.
  
  
  METHODS
  The brain slices (200 µm in thickness) of neonatal SD rats (3 to 5 days old) were prepared and cultured in modified serum-free DMEM/F12 medium at 37 degrees celsius; with 95% O(2) and 5% CO(2). The organotypic slice cultures were observed regularly. NSCs isolated from the cortex of rat embryos (14-15 embryonic days) were cultured in serum-free DMEM/F12 supplemented with B27 and N2, and the passage 3 NSCs were labeled by CM-DiI before transplanted onto the organotypic slices cultured for 2 weeks. The survival of transplanted NSCs was assessed, and the cell differentiation was identified by immunofluorescence staining.
  
  
  RESULTS
  The organotypic slice cultures were well maintained for at least 4 weeks in the modified medium. The thickness of the organotypic slices reduced from 200 µm to 130 µm after 2-week culture in vitro due to the migration of the cells on the edge of the slices. CM-DiI-labeled NSCs survived well and differentiated into GFAP(+) glia and β-tubullin III(+) neurons.
  
  
  CONCLUSIONS
  Neonatal rat organotypic brain slice can be successfully cultured in a modified condition to serve as a model for studying NSC differentiation induced by spatial signals.
authors.affiliation: Institute of Neurobiology, Xi'an Jiaotong University College of Medicine, China. jiaoqian2006@126.com
authors.forename: Qian
authors.initials: Q
authors.lastname: Jiao
authors.forename: Hai-xia
authors.initials: HX
authors.lastname: Zhang
authors.forename: Hai-xia
authors.initials: HX
authors.lastname: Lv
authors.forename: Yong
authors.initials: Y
authors.lastname: Liu
authors.forename: Jin-long
authors.initials: JL
authors.lastname: Li
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002454:Cell Differentiation / Q000502:physiology*; D002540:Cerebral Cortex / Q000166:cytology*; D018920:Coculture Techniques / Q000379:methods; D005333:Fetus; D058953:Neural Stem Cells / Q000166:cytology* / Q000637:transplantation; D009924:Organ Culture Techniques / Q000379:methods*; D051381:Rats