Fig. 2: hEC co-culture increases the percentage of GFAP+LeX+ and GFAP+S100A6+ human type B cells.
From: Human endothelial cells promote a human neural stem cell type B phenotype via Notch signaling
a SC27 hNSPCs differentiated (Diff) for 5 days and immunostained for LeX demonstrate a significant decrease in the percentage of LeX+ hNSPCs compared to undifferentiated hNSPCs in proliferation media (Prolif)(****p < 0.0001). b Co-culture of SC27 hNSPCs with hECs and immunostaining for cell surface markers LeX for hNSPCs and CD31 for hECs show cell contact in confocal imaging (scale bar 30 µm). c SC27 hNSPCs in MC, hEC CM, or with hECs stained for GFAP and LeX. d GFAP+LeX+ type B hNSPCs are significantly greater in co-culture with hECs compared to MC (****p < 0.0001) or hEC CM (****p < 0.0001). Type B hNSPCs out of total number of LeX+ hNSPCs are significantly greater in co-culture with hECs compared to MC (****p < 0.0001) or hEC CM (****p < 0.0001). LeX+ hNSPCs are significantly greater in co-culture with hECs compared to MC (***p = 0.0002) or hEC CM (****p < 0.0001). There was no significant difference between hEC CM and MC for GFAP+LeX+ (p = 0.971), GFAP+LeX+ out of LeX+ (p = 0.478), and LeX+ hNSPCs (p = 0.6614). e HNSPCs differentiated for 21 days and immunostained for S100A6. The percentage of S100A6+ hNSPCs significantly decreased for 5-day (*p = 0.0154) and 21-day (****p < 0.0001) differentiated hNSPCs compared to undifferentiated hNSPCs. f SC27 hNSPCs in MC, hEC CM, or + hECs immunostained for GFAP and S100A6. g The percentage of GFAP+S100A6+ type B hNSPCs is significantly greater in hEC co-culture compared to MC (****p < 0.0001). The percentage of GFAP+S100A6+ hNSPCs out of S100A6+ hNSPCs is significantly greater in hEC co-culture compared to MC (****p < 0.0001) and +hEC CM (****p < 0.0001). The percentage of S100A6+ hNSPCs does not significantly differ in hEC co-culture compared to MC (p = 0.5816) and +hEC CM (0.2749). There was no significant difference between hEC CM and MC for GFAP+S100A6+ (p = 0.0586), GFAP+S100A6+ out of S100A6+ (p = 0.0518), and S100A6+ hNSPCs (p = 0.8424). Co-cultures were grown for 5 days. Nuclei were stained with Hoechst. Scale bars, 100 µm unless otherwise noted, n = 3 independent biological replicates indicated by symbols, 5 areas quantified per sample, percentages are out of total hNSPCs unless otherwise noted, mean with SEM. Analysis used (a, d) unpaired two-tailed Student’s t-test and (c, f) one-way ANOVA with Tukey post-hoc test for multiple comparisons. Source data are provided as a Source Data file.
