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3D artificial round section micro-vessels to investigate endothelial cells under physiological flow conditions

机译:3D人工圆形截面微血管在生理流动条件下研究内皮细胞

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摘要

In the context of xenotransplantation, in ischemia/reperfusion injury as well as in cardiovascular research, the study of the fascinating interplay between endothelial cells (EC) and the plasma cascade systems often requires in vitro models. Blood vessels are hardly reproducible with standard flat-bed culture systems and flow-plate assays are limited in their low surface-to-volume ratio which impedes the study of the anticoagulant properties of the endothelial cells. According to the 3R regulations (reduce, replace and refine animal experimentation) we developed a closed circuit microfluidic in vitro system in which endothelial cells are cultured in 3D round section microchannels and subjected to physiological, pulsatile flow. In this study, a 3D monolayer of porcine aortic EC was perfused with human serum to mimic a xenotransplantation setting. Complement as well as EC activation was assessed in the presence or absence of complement inhibitors showing the versatility of the model for drug testing. Complement activation products as well as E-selectin expression were detected and visualized in situ by high resolution confocal microscopy. Furthermore, porcine pro-inflammatory cytokines as well as soluble complement components in the recirculating fluid phase were detected after human serum perfusion providing a better overview of the artificial vascular environment.
机译:在异种移植的背景下,在缺血/再灌注损伤以及心血管研究中,对内皮细胞(EC)和血浆级联系统之间令人着迷的相互作用的研究通常需要体外模型。用标准的平板培养系统很难重现血管,流板法的低表面积体积比受到限制,这妨碍了对内皮细胞抗凝特性的研究。根据3R法规(减少,替换和完善动物实验),我们开发了一种闭路微流体体外系统,其中内皮细胞在3D圆形截面微通道中培养,并经受生理性脉动流。在这项研究中,用人血清灌注猪主动脉EC的3D单层膜以模拟异种移植设置。在存在或不存在补体抑制剂的情况下评估补体以及EC激活,显示出药物测试模型的多功能性。补体激活产物以及E-选择蛋白表达被检测到并通过高分辨率共聚焦显微镜原位显示。此外,在人血清灌注后,可检测到猪促炎细胞因子以及循环液中的可溶性补体成分,从而更好地了解了人工血管环境。

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