Figure 5.

Analysis of the hybrid approach based on whole mount microvessel properties. (A-C) Representative images of thick longitudinal sections of slabs using the hybrid approach, in which some slabs were harvested at day 3 (A) and others were cultured for an additional 3 days in either defined medium (B) or EGM-2+ (C). HUVECs were stained for CD31 (red), PCs were GFP-labeled (green) and nuclei were stained blue. The long axes of the slabs are vertical. Scalebars = 100 μm. (D-G) Macroscopic images (D,F) and alignment maps (E,G) of slabs cultured in defined medium (D-E) or EGM-2+ (F-G). The red segments in the alignment maps indicate the local strength and direction of fibril alignment. (H-J) Alignment and compaction parameters indicated that the slabs cultured in EGM-2+ had higher fibrin fibril alignment (birefringence; H), compacted more (smallest width at the middle of the slab; I), and had higher microvessel alignment (anisotropy index; J) than slabs cultured in defined medium. (K-M) Microvascular network characteristics, including total microvessel length per area (K) and average structure length (L), were similar between slabs cultured in EGM-2+ and the control slabs, but reduced in slabs cultured in defined medium. PC recruitment (M) was similar across all conditions. *p < 0.05.