Figure 13.

Effect of phagocytic microglia secreted factors on late neurogenesis in vitro. A, Experimental design of the in vitro late survival and differentiation assay. B, Density of live and dead cells found after 10 d of DMEM/F12 followed by 3–5 d of CM microC or microPH and DMEM. The number of cells before adding the CM (t = 0) is shown as a control. C, Representative confocal microscopy images of NPCs treated for 10d with DMEM/F12 followed by 3–5 d of CM microC, microPH, or DMEM. Top, Left, DMEM/F12 treatment of 10 d, before adding any CM. D, Percentage of cell types found after 10 d of DMEM/F12 followed by 3–5 d of CM microC or microPH and DMEM. The number of cells before adding the CM (t = 0) is shown as a control. Mat stellate designates stellate cells with mature (more branched) morphology, and early/late ram designates early/late ramified cells. Scale bars: 20 μm, z = 6.3 μm. N = 3 independent experiments (B, D). Three-way ANOVA (treatment × life × time, F) showed interactions between the different factors and thus the data were split into several one-way ANOVAs, which showed no significant effect of the treatment. Data in D could not be normalized because some cell categories were only present in particular treatments (i.e., the mature stellate phenotype only occurred in MicroPH groups). Error bars represent mean ± SEM. Values of statistics used are shown in Table 8.