Smith Ashley M, Prabhakarpandian Balabhaskar, Pant Kapil
Biomedical Technology, CFD Research Corporation.
Biomedical Technology, CFD Research Corporation;
J Vis Exp. 2014 May 25(87):51025. doi: 10.3791/51025.
Cell/particle adhesion assays are critical to understanding the biochemical interactions involved in disease pathophysiology and have important applications in the quest for the development of novel therapeutics. Assays using static conditions fail to capture the dependence of adhesion on shear, limiting their correlation with in vivo environment. Parallel plate flow chambers that quantify adhesion under physiological fluid flow need multiple experiments for the generation of a shear adhesion map. In addition, they do not represent the in vivo scale and morphology and require large volumes (~ml) of reagents for experiments. In this study, we demonstrate the generation of shear adhesion map from a single experiment using a microvascular network based microfluidic device, SynVivo-SMN. This device recreates the complex in vivo vasculature including geometric scale, morphological elements, flow features and cellular interactions in an in vitro format, thereby providing a biologically realistic environment for basic and applied research in cellular behavior, drug delivery, and drug discovery. The assay was demonstrated by studying the interaction of the 2 µm biotin-coated particles with avidin-coated surfaces of the microchip. The entire range of shear observed in the microvasculature is obtained in a single assay enabling adhesion vs. shear map for the particles under physiological conditions.
细胞/颗粒黏附测定对于理解疾病病理生理学中涉及的生化相互作用至关重要,并且在寻求新型治疗方法的开发中具有重要应用。使用静态条件的测定无法捕捉黏附对剪切力的依赖性,限制了它们与体内环境的相关性。在生理流体流动下量化黏附的平行板流动腔需要多次实验才能生成剪切黏附图。此外,它们不能代表体内的尺度和形态,并且实验需要大量(约毫升)的试剂。在本研究中,我们展示了使用基于微血管网络的微流控装置SynVivo-SMN从单个实验中生成剪切黏附图。该装置以体外形式重现了复杂的体内脉管系统,包括几何尺度、形态学元素、流动特征和细胞相互作用,从而为细胞行为、药物递送和药物发现的基础研究和应用研究提供了一个生物学上逼真的环境。通过研究2微米生物素包被颗粒与微芯片抗生物素蛋白包被表面的相互作用来证明该测定。在单个测定中获得了在微脉管系统中观察到的整个剪切力范围,从而能够得到生理条件下颗粒的黏附与剪切力图谱。