Fig. 2: 3D culture models kidney morphogenesis dynamics more closely than ALI cultures.
From: Engineering kidney developmental trajectory using culture boundary conditions
A Top, average immunofluorescence projection of E12 kidney in 3D 0.5 C + M culture. Representative of 11 kidneys from 3 experiments. Bottom, UB outlines over time. Both are color-coded by culture time. B Schematic and plot of E13 kidney UB tip distance after culture in ALI or 3D formats for 3 days, compared to size-matched E14 kidney (n = 21 (ALI), 24 (3D) and 34 (E14) tip pairs, 3 kidneys each, pooled across 3 litters. One-way ANOVA with Tukey’s test, p = 0.0002 and 0.0016 for ALI vs 3D culture and ALI vs E14.) C Schematic of dihedral angle change during branching. D Schematic of physics-based model definition. E Simulation and quantification of branching of UB trees under physical constraints that model ALI (Left) vs. 3D hydrogel (Middle) cultures. Left and middle, perspective and top views of the UB branch families before and after energy minimization within the bounding box (gray). Right, quantification of branching angle change of left and middle. (n = 24 and for both ϕ and θ; unpaired two-tailed t-tests, p = 2.63*10−13). F Live images tracking E13 kidney UB tips over 11 h in ALI vs. 3D cultures. Insets show tips (yellow circles) and tip position tracks (red/brown lines) for the indicated time points. G Quantification of track rotation velocity in (F). (n = 45 from 3 E12 and 1 E13 kidneys for ALI culture and 75 tips from 4 E12 and 2 E13 kidneys for 3D culture, pooled across 2 and 3 litters, respectively; unpaired two-tailed t-tests, p = 4.08*10−22). All plotted as mean ± S.D. **p < 0.01, ***p < 0.001, ****p < 0.0001.
