Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation

Figure 4

NPCs in 3D neurospheres have increased homogenous PAX6 expression. (a) Flow cytometer dot plot demonstrated that PAX6 expression in 3D NPCs was more homogeneous compared to 2D NPCs. (b) Specific gating was applied to quantify the low PAX6 expression population in 2D NPCs and 3D NPCs. Student’s -test ( and ). The bars represent the Mean ± SEM of 3 independent cultures set. (c) QPCR analysis of SOX2, NESTIN, and PAX6 expression of cultured 2D NPCs and 3D NPCs. Expression values were normalized to GAPDH (reference gene). Subsequently, the expression values were calculated as relative amount of mRNA versus expression values of N S5 (monolayer) which was set to 1. Test for significant difference () using Student’s -test. The bars represent the Mean ± SEM of 3 independent cultures set. (d) A scatter plot and trend line (Pearson correlation) showed correlation between day 7 neurosphere diameter and the average relative amount of mRNA of PAX6 in 3D NPCs.