Research Article

Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation

Figure 5

Neurosphere aggregation promotes astrocyte differentiation. (a) ICC of 21-day differentiated neurons and astrocytes (TUBB3 and GFAP) derived from 2D and 3D NPCs. (b) ICC of 21-day neurons (MAP2 and TUBB3) differentiated from 2D and 3D NPCs. (c) Quantification of TUBB3+ and GFAP+ cells from iPSC-derived NPCs. Results were reported as Mean ± SEM of 15 fields from 3 independent cultures, tested for significant difference ( and ) using Student’s -test. (d) Gene expression by qPCR analysis of TUBB3, MAP2, and GFAP. The expression values were normalized to GAPDH. Subsequently, the expression values were calculated as relative amount of mRNA versus expression values of N S5 (derived from 2D NPCs) 21-day differentiated cells, which was set to 1. Data was reported as Mean ± SEM of 3 independent cultures, tested for significant difference ( and ) using Student’s -test. (e) A scatter plot and trend line (Pearson correlation) showed correlation between the average relative amount of GFAP (21-day differentiated cells) and BLBP (NPCs) mRNA. The expression values of BLBP in NPCs were calculated as relative amount of mRNA versus expression values of N S5 (monolayer) which was set to 1. Data was obtained from 3 independent cultures. (f) A scatter plot and trend line (Pearson correlation) showed correlation between the average relative amount of GFAP (21-day differentiated cells) and PAX6 (NPCs) mRNA.