Fig. 4

Proliferation, differentiation, and immunofluorescent staining of neural stem cells (NSCs) encapsulated in the hydrogels. A Cell proliferation of NSCs embedded in the CO and COA2 hydrogels measured by CCK-8 assay during a culture period of 14 days. The proliferation was normalized to the value at day 0 and expressed as the percentage of the cell proliferation (%). B Expression of neural-related genes in NSCs encapsulated in the hydrogels were analyzed by RT-PCR at 14 days. The expression levels normalized to that of housekeeping gene (GAPDH) and represented by the relative ratios of gene expression. Immunofluorescent images of NSCs stained for (C) GFAP and (D) MAP2 at 14 days. Scale bars represent for 100 μm. E Quantification of the fluorescent intensity in GFAP- and MAP2-positive NSCs. *p < 0.05, **p < 0.01, and ***p < 0.001 between the indicated groups