Establishment of an adult neural stem cell in vitro model of ischemia. (a) The identification of adult neural stem cells throughout different passages. Immunocytochemical detection of nestin (red) was performed. The nuclei of NSCs were revealed by DAPI staining (blue). The scale bar represents 20 μm. (b) Digital photomicrographs of NSCs exposed to different durations of in vitro ischemia. NSCs were subjected to OGD for different periods (0–10 h), then were returned to normoxic conditions and incubated for an additional 24 h. The cells were photographed at the end of the experimental period. All photomicrographs are from different sister cultures from the same plating. Minor adjustments to brightness, contrast and color balance have been made to the digital images. The scale bar represents 20 μm. (c) Cell viability and survival rate of the NSCs following in vitro ischemia. To estimate the number of viable cells, approximately 50,000 cells were grown in each well of poly-L-lysine-coated 96-well plates with 100 μL medium and the absorbance at 490 nm was directly proportional to the number of viable NSCs per well at each time point. Data points represent the mean ± SD of six independent experiments. *P < 0.05 versus control, analyzed by one-way ANOVA/Bonferroni post hoc test.
Chen et al. BMC Neuroscience 2013 14:24 doi:10.1186/1471-2202-14-24