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Merck
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  • Design and Validation of a Human Brain Endothelial Microvessel-on-a-Chip Open Microfluidic Model Enabling Advanced Optical Imaging.

Design and Validation of a Human Brain Endothelial Microvessel-on-a-Chip Open Microfluidic Model Enabling Advanced Optical Imaging.

Frontiers in bioengineering and biotechnology (2020-10-30)
Mootaz M Salman, Graham Marsh, Ilja Kusters, Matthieu Delincé, Giuseppe Di Caprio, Srigokul Upadhyayula, Giovanni de Nola, Ronan Hunt, Kazuka G Ohashi, Taylor Gray, Fumitaka Shimizu, Yasuteru Sano, Takashi Kanda, Birgit Obermeier, Tom Kirchhausen
摘要

We describe here the design and implementation of an in vitro microvascular open model system using human brain microvascular endothelial cells. The design has several advantages over other traditional closed microfluidic platforms: (1) it enables controlled unidirectional flow of media at physiological rates to support vascular function, (2) it allows for very small volumes which makes the device ideal for studies involving biotherapeutics, (3) it is amenable for multiple high resolution imaging modalities such as transmission electron microscopy (TEM), 3D live fluorescence imaging using traditional spinning disk confocal microscopy, and advanced lattice light sheet microscopy (LLSM). Importantly, we miniaturized the design, so it can fit within the physical constraints of LLSM, with the objective to study physiology in live cells at subcellular level. We validated barrier function of our brain microvessel-on-a-chip by measuring permeability of fluorescent dextran and a human monoclonal antibody. One potential application is to investigate mechanisms of transcytosis across the brain microvessel-like barrier of fluorescently-tagged biologics, viruses or nanoparticles.

材料
Product Number
品牌
产品描述

Sigma-Aldrich
Pluronic® F-127, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
胶原蛋白 来源于人类胎盘, Bornstein and Traub Type IV, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
哌嗪-N,N′-二(2-乙磺酸), ≥99% (titration)
Sigma-Aldrich
京尼平, ≥98% (HPLC), powder
Sigma-Aldrich
异硫氰酸荧光素-葡聚糖, average mol wt 10,000
Sigma-Aldrich
铁氰化钾 (II) 三水合物, ≥99.95% trace metals basis
Sigma-Aldrich
聚二甲基硅氧烷, viscosity 1.0 cSt (25 °C)
Sigma-Aldrich
双酚A型环氧树脂, used as embedding medium
Sigma-Aldrich
纤连蛋白 人类包皮成纤维细胞, lyophilized powder, suitable for cell culture