Fig. 3

Establishment of iPSC-derived neurons and astrocytes co-culture system and cellular changes of the iPSC-derived astrocytes in the co-culture system. a Schematic experimental paradigm of the co-culture using individually induced neurons and astrocytes from the same iPSCs. b Bright-field images of the co-culture model of the iPSC-derived neurons (white arrow) and astrocytes (yellow arrow) at 30 days after co-culturing of the cells. Scale bar; 100 μm. c Representative immunocytochemistry images of the iPSC-derived mono-culture model of astrocytes and co-culture model, at 30 days after co-culturing of the cells, stained with antibodies specific for S100β, GFAP, and MAP2. Scale bar; 100 μm. d Representative images of iPSC-derived astrocytes in the mono- and co-culture models. e Sholl analysis showing the complexity of the iPSC-derived astrocyte of the mono-culture model (n = 30) and co-culture model (n = 30) (three independent experiments). Bars, mean ± SEM. ****p < 0.0001 (Two-way ANOVA). f Immunocytochemistry images of iPSC-derived astrocytes stained with S100β and GFAP in the mono- and co-culture models. Scale bar; 100 μm. g Quantification and comparison of the GFAP/S100β ratios between the mono-culture model (n = 45) and the co-culture model (n = 45). ****p < 0.0001 (Mann-Whitney test)