Abstract
We present a novel methodology based on ion conductance to evaluate the perfusability of vascular vessels in microfluidic devices without microscopic imaging. The devices consisted of five channels, with the center channel filled with fibrin/collagen gel containing human umbilical vein endothelial cells (HUVECs). Fibroblasts were cultured in the other channels to improve the vascular network formation. To form vessel structures bridging the center channel, HUVEC monolayers were prepared on both side walls of the gel. During the culture, the HUVECs migrated from the monolayer and connected to the HUVECs in the gel, and vascular vessels formed, resulting in successful perfusion between the channels after culturing for 3–5 d. To evaluate perfusion without microscopic imaging, Ag/AgCl wires were inserted into the channels, and ion currents were obtained to measure the ion conductance between the channels separated by the HUVEC monolayers. As the HUVEC monolayers blocked the ion current flow, the ion currents were low before vessel formation. In contrast, ion currents increased after vessel formation because of creation of ion current paths. Thus, the observed ion currents were correlated with the perfusability of the vessels, indicating that they can be used as indicators of perfusion during vessel formation in microfluidic devices. The developed methodology will be used for drug screening using organs-on-a-chip containing vascular vessels.
Highlights
Vascular organs play a crucial role in the transport of nutrients and cancer metastasis, and vessel formation has been widely investigated in vitro
Such devices provide perusable capillary networks consisting of vascular endothelial cells [5], with blood vessels formed by vasculogenesis and angiogenesis [6]
The gel tobetween induce vascular anastomosis on angiogenesis andincorporated vasculogenesis the In geladdition, to inducehLFs vascular anastomosis based on angiogenesis and vasculogenesis
Summary
Vascular organs play a crucial role in the transport of nutrients and cancer metastasis, and vessel formation has been widely investigated in vitro. To mimic in vivo environments, vascular endothelial cells are typically co-cultured with fibroblasts both two- and threedimensionally [3,4], resulting in the successful formation of capillary networks with lumen structures. Vascular models have been constructed in microfluidic devices to resemble in vivo environments and to arrange cells/extracellular matrices (ECMs). Such devices provide perusable capillary networks consisting of vascular endothelial cells [5], with blood vessels formed by vasculogenesis and angiogenesis [6]. The formation of new blood vessels, known as vasculogenesis, occurs early in the developmental stage of a vascular tissue; progenitor cells rearrange themselves to form a lumen, resulting in the formation of new capillary blood vessels. New blood vessels form from pre-existing vessels through vascular sprouting
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