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Development and Characterization of In Vitro Microvessel Network and Quantitative Measurements of Endothelial [Ca2+]i and Nitric Oxide Production
Published on: May 19, 2016
Vascular lumen simulation and highly-sensitive nitric oxide detection using three-dimensional gelatin chip coupled to
Lin-Mei Li1, Xue-Ying Wang, Liang-Sheng Hu
1Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, 430072, China.
Abstract:
Reproducing the physiological environment of blood vessels for the in vitro investigation of endothelial cell functions is very challenging. Here, we describe a vascular-like structure based on a three-dimensional (3D) gelatin chip with good compatibility and permeability which is also cost-effective and easy to produce. The controllable lumen diameter and wall thickness enable close mimicking of blood vessels in vitro. The 3D gelatin matrix between adjacent lumens is capable of generating soluble-factor gradients inside, and diffusion of molecules with different molecular weights through the matrix is studied. The cultured human umbilical vein endothelial cells proliferate on the gelatin lumen linings to form a vascular lumen. The hemodynamic behavior including adhesion, alignment of endothelial cells (ECs) under shear stress and pulsatile stretch is studied. Furthermore, a microelectrode comprising TiC/C nanowire arrays is fabricated to detect nitric oxide with sub-nM detection limits and NO generation from the cultured ECs is monitored in real time. This vascular model reproduces the surrounding parenchyma of endothelial cells and mimics the hemodynamics inside blood vessels very well, thereby enabling potential direct investigation of hemodynamics, angiogenesis, and tumor metastasis in vitro.

