Fig. 4
In vitro vessel-like network maturity. a Representative confocal images of whole-mount immunofluorescent scaffolds populated with tri-cultures. EC endothelial cells, SMC smooth muscle cells. Endothelial cell-ZsGreen are stained in green, collagen IV-positive cells are stained in red and nuclei are stained in blue; scale bar = 100 µm. b Representative confocal images of whole-mount immunofluorescent scaffold populated with endothelial cellANGPT1-SMCVEGF-myoblast tri-culture. Endothelial cellANGPT1-ZsGreen are stained in green, collagen IV-positive cells are stained in red and nuclei are stained in blue; scale bar = 10 µm. c Representative confocal images of whole-mount immunofluorescent scaffolds populated with co- or tri-cultures. Endothelial cell-ZsGreen are stained in green, VE-cadherin-positive cells are stained in red and nuclei are stained in blue; scale bar = 10 µm. d Collagen IV-wrapped vessels. Myo myoblasts. Data are expressed as box-and-whisker plots, where the central lines denote medians, edges represent upper and lower quartiles and whiskers show minimum and maximum values. Data were analyzed by one-way ANOVA, followed by Holm–Sidak’s multiple comparison test, n = 5 (*p = 0.0176 for ECANGPT1-SMC versus EC-SMCs co-culture, *p = 0.0326 for ECANGPT1-SMCVEGF versus EC-SMC co-culture, **p = 0.0077 for EC-SMCVEGF versus EC-SMC co-culture, ***p = 0.0001 for ECANGPT1-SMCVEGF-Myo versus EC-SMC co-culture, ****p < 0.0001 for EC-SMC-Myo versus EC-SMC co-culture). e VE-cadherin-positive vessel elongation. Data are expressed as box-and-whisker plots, where the central lines denote medians, edges represent upper and lower quartiles and whiskers show minimum and maximum values. Data were analyzed by one-way ANOVA, followed by Holm–Sidak’s multiple comparison test, n = 5 (*p = 0.0119 for ECANGPT1-SMC versus EC-SMC co-culture, **p = 0.0015 for EC-SMCVEGF versus EC-SMC co-culture, **p = 0.0051 for ECANGPT1-SMCVEGF-Myo versus EC-SMC co-culture, ***p = 0.0004 for EC-SMC-Myo versus EC-SMC co-culture)
