Fig. 10.

Intrauterine exposure to gestational diabetes mellitus (GDM) alters endothelial colony-forming cell (ECFC) response to laminar flow. A: ECFCs from uncomplicated (UC) and GDM pregnancies were plated on type 1 collagen-coated glass slides and were maintained in static culture or exposed to 7 h of laminar flow. White arrows in the top right of image panels indicate flow directionality. Following treatment, ECFCs were fixed and stained for transgelin (TAGLN) (red), F-actin (green), and NucBlue (blue). All 3 channels were overlaid in the merged image. Scale bars, 25 μm. B: ECFC alignment was analyzed with the Directionality Plug-in (FIJI). The F-actin fluorescence images were used to produce the histograms representing the distribution of angles measured within 8 images for each sample, which were combined and overlaid for a total of 200–250 cells analyzed per sample. C: box and whisker plots representing statistical analysis completed to compare angle variance distributions between the 2 groups (UC and GDM) in the presence and absence of flow (UC: n = 5; GDM: n = 8). Data were analyzed by two-way ANOVA. A significant interaction was identified between GDM exposure and laminar flow (P = 0.028). Angle variance was significantly altered by laminar flow exposure (P < 0.0001).