Figure 5
From: Engineered 3D vascular and neuronal networks in a microfluidic platform
hiPS-vascular and hES-neuronal networks in a microfluidic device. (a,b,c) Formation of iPS-EC networks in a microfluidic device with MN spheroids. Microvascular networks were formed in 2–3 days. After formation of the microvascular network, MN spheroids start to extend protrusions and form neuronal networks intermingling with the microvascular networks. Magnified region is indicated in the white dotted box. (d,i) Lumen formation of iPS-EC vascular networks. The average diameter of networks is approximately 55 μm with MN networks, while the diameter is less than 40 μm without MN networks. n = 6; *P < 0.01, student’s t-test. (e) Evaluation of vascular connectivity by perfusion of 1 μm microbeads through microvascular networks. Images show the path lines of microbeads over a 9 second period. (f) Evaluation of neuronal connectivity by Ca2+ imaging stained by fluo-8-AM. Frequent Ca2+ oscillation can be observed in microfluidic devices. (g,h) Quantification of Ca2+ oscillation with and without iPS-EC networks. Local Ca2+ transportation was analyzed by post-image processing. (g) Frequency and amplitude of Ca2+ oscillation increased in the presence of iPS-EC networks. (j) Relative gene expression change between with and without EC networks after 7 days of static culture indicated that iPS-EC microvascular networks clearly upregulated the gene related to MN differentiation, neurite outgrowth, and synaptic formation. n = 3.
