Figure 8.

Development and characterization of the segregated NV platform. (A) Illustration of the platform and assembly method via integration of two separate components: a 3D vessel network platform (top) and a 3D neural tissue platform (bottom), both at 11 DIV. The segregated NV platform is cultured for further 10 DIV in the 1:1 medium (see the Methods section for further details). Illustration made with biorender (https://biorender.com/). (B) Vessel morphology (shown by CD31 in red) in the NV platform at 21 DIV (bottom) shows further maturation denoted by thinning, less branching, and a straighter pattern compared to vessels at 11 DIV from the vascular-only model (top). Scale bars represent 100 μm. (C) Vascular development in the vicinity of the neurosphere, within the segregated NV platform (21 DIV). HMVECs (immunostained by CD31, red) were attracted to the neurosphere (immunostained by βIII-tubulin, white) and formed either a monolayer when in direct contact (top row) or a vascular plexus when not in direct contact (bottom row). Right, merged images. Nuclei stained with DAPI (blue). Scale bars represent 100 μm and apply to the panels in the respective row. (D) Vessel innervation occurred primarily at regions farther away from the neurosphere cluster, where neurites (βIII-tubulin, white) could innervate and align with the vessels (CD31, red) as highlighted in the yellow-dashed box, zoomed in the top right corner box. Scale bar is 50 μm.