3-dimensional cytoarchitecture of vasculature-like structures.

<p>(A) A 3-dimensional cytoarchitecture composed of hCMECs (CD31+ cells in red) and hNSCs (polyclonal GFAP+ cells in green) was visualized using confocal microscopy. (B) At the base of the coculture, hNSCs were seen in a monolayer on which hCMECs formed VLS that was characterized by a multicellular organization that extended beyond the monolayer to form a unique structure. (C) Some hNSCs with an astrocytic phenotype were associated with these VLS by forming a layer of cells around the tubular walls in some cases with an elongated morphology and endfeet on the VLS (yellow arrows). The astrocytes were ensheathing the VLS. (D) A higher magnification 3D confocal image clearly demonstrates structural differences in astrocytes’ morphology upon interfacing with the VLS rather than being inside the neural patch. (E) A 3-dimensional cut through this area further indicates that astrocytes provide structural support (white arrows) to the VLS. The orange line indicates the monolayer formed by hNSCs onto which the VLS rests. Nevertheless, no hollow lumen is formed by the VLS. It hence here remains unclear if a 3-dimensional support structure is required to create a lumen and if they are inflatable in a flow system. Diamidino-2-phenylindole (DAPI, blue) serves as a nuclear counterstain. Scale bars represent 100 µm.