Figure 4

In vitro replication of cerebellar anlage at the maturation stage. NPCs cultured in NS21 medium matured to calbindin-positive PNs. (a) Culture of NPCs in NS21 medium comprised calbindin-positive and GFAP-positive cells. (b) Calbindin-positive and NeuN-positive cells were present after 30 DIV in NS21 medium. There was co-expression of (c) calbindin and parvalbumin, and (d) calbindin and IP3R in NPCs cultured in NS21 medium. NPCs from 11 to 15 DIV EBs in SFEBq culture presented various cell types in culture and are shown as ratios of: (e) calbindin-positive cells to GFAP-positive cells; (f) GFAP-positive cells with respect to the total DAPI-positive cells; (g) calbindin-positive cells with respect to the total DAPI-positive cells; and (h) NeuN-positive cells to the total DAPI-positive cells. Data are represented as the mean and SEM (n = 10 images from 6 experimental runs consisting of 6 cell batches), which were assessed via one-way ANOVA followed by Tukey’s with statistical significance set at P ≤ 0.01. Scale bars: 60 μm (a); 200 μm (b); 20 μm (c and d).