Figure 2

Characterization of neural stem cell-like cells (iNSCs) induced from hADSCs by the optimized induction method. (a) Neurosphere formation of iNSCs in suspension at DIV 4 after passage. Scale bar: 100 μm. (b) Immunocytochemistry analysis of the expressions of Nestin and SOX2 in iNSCs and hADSCs cultured as a monolayer on cover slips. Scale bar: 20 μm. (c) Real-time qPCR of NSC and early neuronal markers, Nestin, Sox1, Sox2, Pax6, Musashi-1, Vimentin, Emx1, Gli3, FoxG1, Nkx2.1, Dlx2, Ascl1, Tuj1 and Olig2, at each step of the induction process. The mRNA level of a given gene was quantified by densitometry and normalized to the corresponding GAPDH level. The bars represent the mean ± SEM of at least three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 compared to hADSC, ^†P < 0.05, ^††P < 0.01, ^†††P < 0.001 compared to STEP 1, ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 compared to STEP 2. ANOVA followed by post hoc Newman-Keuls test. S1, STEP 1; S2, STEP 2; S3, STEP 3. (d) Transplantation of iNSCs onto the ventral horn of rat organotypic spinal cord slices. The transplanted iNSCs were stained with monoclonal anti-human nuclei (hNu, red) and either polyclonal anti-SOX2 or anti-TuJ1 antibodies (green). Scale bar: 100 μm.