Organotypic slice culture of neonatal rat cortex and induced neural stem cell differentiation / 南方医科大学学报
Journal of Southern Medical University
; (12): 1318-1322, 2011.
Article
in Zh
| WPRIM
| ID: wpr-235134
Responsible library:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>
Full text:
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Index:
WPRIM
Main subject:
Organ Culture Techniques
/
Physiology
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Transplantation
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Cell Differentiation
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Cerebral Cortex
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Coculture Techniques
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Cell Biology
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Neural Stem Cells
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Fetus
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Animals, Newborn
Limits:
Animals
Language:
Zh
Journal:
Journal of Southern Medical University
Year:
2011
Type:
Article