Organotypic Cocultures of Human Pluripotent Stem Cell Derived-Neurons with Mammalian Inner Ear Hair Cells and Cochlear Nucleus Slices

Figure 5

The number of VGLUT1-positive puncta was greater in CN-SC cocultures compared with CN slice controls. The number and size of VGLUT1 puncta in immunofluorescence images from CN-only controls (a) and CN-SC cocultures (b) at 14 DIV were analysed using ImageJ Analyze Particles function (c, d; ImageJ drawings of VGLUT1 puncta above a threshold). The number of VGLUT1-positive puncta (normalised to percentage of neurofilament-positive area in frame) was compared between CN-only controls and CN-SC cocultures. When independent two-sample -tests were performed, the number of puncta was significantly higher compared with CN slice controls (e, , independent samples -test with 95% confidence interval). Conversely, the average size of VGLUT1-positive puncta between the two groups did not differ significantly (ns) in these sample sizes (f, -test: mean difference of -0.0015, 95% CI of -0.0035, 0.0005, ). Scale bars in (b) and (d) = 20 μm, applies to panels (a) and (c), respectively.