Fig. 2: Deposition of mature glomerular extracellular matrix proteins in GlomSpheres, compared to podocyte and GEnC mono-culture spheroids.
From: GlomSpheres as a 3D co-culture spheroid model of the kidney glomerulus for rapid drug-screening
a Podocyte only spheroid stained for collagen IV α1, which is expressed. b Podocyte only spheroid stained for collagen IV α3, which is not strongly expressed. c Podocyte only spheroid stained for laminin α5, which is not strongly expressed. d GEnC only spheroid stained for collagen IV α1, which is expressed. e Endothelial only spheroid stained for collagen IV α3, which is not strongly expressed. f GEnC only spheroid stained for laminin α5, which is not strongly expressed. g Co-cultured GlomSphere stained for collagen IV α1, which is expressed. h Co-cultured GlomSphere stained for collagen IV α3, which has increased expression compared to monocultured spheroids. i Co-cultured GlomSphere stained for laminin α5, which is stained more strongly than monocultured spheroids. j Mean fluorescence intensity quantification for collagen IV α1 staining (minimum n = 16). Mean fluorescence intensity is not significantly different between conditions (one-way ANOVA p = 0.0814). k Mean fluorescence intensity quantification for collagen IV α3 staining (minimum n = 19). GEnC expression was found to be significantly higher than podocytes (Tukey’s multiple comparison *P = 0.0408). Co-cultured (GlomSphere) expression is shown to be significantly higher than both Podocytes (****P = < 0.0001) and GEnCs (****P = < 0.0001). l Mean fluorescence intensity quantification for laminin α5 staining (minimum n = 7). Podocyte and GEnC expression are shown to be insignificantly different (Tukey’s multiple comparison *P = 0.4044). Co-culture (GlomSphere) expression is significantly higher than Podocytes (****P = < 0.0001) and GEnCs (****P = < 0.0001). m Western blot data for collagen IV α1 expression. Densitometry (minimum n = 3) showed no significant difference between podocytes, GEnCs and co-culture (one-way ANOVA P = 0.2209). n Western blot data for collagen IV α3 expression. Densitometry (minimum n = 3) showed no significant difference between podocytes and GEnCs (P = 0.9985) Co-culture expression was shown to be significantly higher than both podocytes (****P = < 0.0001) and GEnCs.
