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. 2023 Oct 9;11:1251195. doi: 10.3389/fbioe.2023.1251195

FIGURE 1.

FIGURE 1

Design of the 3D-AD Microfluidic Device and Experimental Setup. (A) 3D rendered image of the microfluidic device consisting of a main BBB chamber with two side channels and an additional neurosphere compartment located in the center of the BBB chamber. Dimensions are indicated in white. (B) Phase contrast image of the BBB cells and neurospheres seeded in the device (Figure 3A bottom). (C) Schematic illustration of the microfluidic device. The suspended cells are injected through the BBB loading port; BBB media is added to the two media channels flanking the central chamber. The neurospheres are seeded into the central neurosphere compartment and the neuronal medium is added from the top. (D) Illustration of the co-culture system. ReN-AD neurospheres seeded in the central compartment secrete factors (e.g., Aβ) that induce pathological changes in the BBB microvascular network formed with primary human microvascular endothelial cells, primary human astrocytes, and primary human microvascular pericytes. (E) Timeline of the experiment: neurospheres are generated 21 days before initiating co-culture with the BBB on day 0. A pressure gradient is initiated on day 2 and the system is analyzed on day 7. Created with BioRender.com.