Figure 1

Soft surfaces increase astrocytes differentiated from mouse embryonic neural stem cells in the absence of serum. (a) Mouse embryonic NSCs were cultured on three types of plate (1 kPa plates, 12 kPa plates, and commonly used plastic plates [2.8 GPa]) for 3 days in serum-free condition and assessed by immunofluorescence staining. Neurons were visualized by Tuj1 (green), astrocytes were visualized by GFAP (red), and cell nuclei were counterstained with Hoechst (blue). Scale bar, 100 μm. (b) Whole cell number, (c) the number of Tuj1 positive cells, and (d) the number of GFAP positive cells on each plate were counted and normalized relative to the control. (e) Protein expression of Tuj1, GFAP, and GAPDH (for loading control) on each plate was detected by western blotting analysis. The expression levels of (f) Tuj1 and (g) GFAP proteins were normalized relative to GAPDH protein. Error bars represent the standard deviation; *p < 0.05, **p < 0.01, ***p < 0.005 (Student’s t test with Bonferroni correction). NSCs neural stem cells, GFAP glial fibrillary acidic protein, GAPDH glyceraldehyde 3-phosphate dehydrogenase.