Upscaling of a living membrane for bioartificial kidney device
Upscaling of a living membrane for bioartificial kidney device
复制标题
DOI：
10.1016/j.ejphar.2016.07.009
复制
发表时间：
2016-11-05
影响因子：
5
通讯作者：
Stamatialis, Dimitrios
中科院分区：
文献类型：
--
作者：
Chevtchik, Natalia Vladimirovna;Fedecostante, Michele;Stamatialis, Dimitrios
透析肾患者代谢废物的清除有限，导致高发病率和死亡率。生物人工肾装置（巴克）可能提供一种更完全去除这些代谢物的强大解决方案，该装置包含具有功能性近端小管上皮细胞（PTEC）的混合“活膜”。这些细胞由人工功能化中空纤维膜（HFM）支撑，能够主动去除废物。在我们早期的研究中，条件永生化人PTEC（ciPTEC）显示当接种在小尺寸的平坦或中空纤维聚醚砜（PES）膜上时表达功能性有机阳离子转运蛋白2（OCT 2）。在这里，提出了一种升级的“活膜”。我们开发并评估了包含三种市售MicroPES HFM支持ciPTEC的模块的功能。HFM最佳地用L-多巴和胶原IV包被以支持在静态培养条件下成熟的ciPTEC的均匀且紧密的单层形成。封闭小带-1（ZO-1）蛋白的丰富表达和FITC-菊粉的有限扩散都证实了单层细胞具有明显的屏障功能。此外，在不存在和存在已知OCT抑制剂（如西咪替丁和阳离子尿毒症溶质混合物）的情况下，研究了荧光OCT 2底物4-（4-（二甲基氨基）苯乙烯基）-N-甲基碘化吡啶（ASP（+））的摄取。在任一抑制剂存在下，活性放大膜的ASP（+）摄取降低了60%，证明了OCT 2的活性功能。总之，本研究提出了一个成功的放大活性有机阳离子传输膜作为支持巴克设备。(C)© 2016 Elsevier B. V.版权所有。
The limited removal of metabolic waste products in dialyzed kidney patients leads to high morbidity and mortality. One powerful solution for a more complete removal of those metabolites might be offered by a bioartificial kidney device (BAK), which contains a hybrid "living membrane" with functional proximal tubule epithelial cells (PTEC). These cells are supported by an artificial functionalized hollow fiber membrane (HFM) and are able to actively remove the waste products. In our earlier studies, conditionally immortalized human PTEC (ciPTEC) showed to express functional organic cationic transporter 2 (OCT2) when seeded on small size flat or hollow fiber polyethersulfone (PES) membranes. Here, an upscaled "living membrane" is presented. We developed and assessed the functionality of modules containing three commercially available MicroPES HFM supporting ciPTEC. The HFM were optimally coated with L-Dopa and collagen IV to support a uniform and tight monolayer formation of matured ciPTEC under static culturing conditions. Both abundant expression of zonula occludens-1 (ZO-1) protein and limited diffusion of FITC-inulin confirm a clear barrier function of the monolayer. Furthermore, the uptake of 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (ASP(+)), a fluorescent OCT2 substrate, was studied in absence and presence of known OCT inhibitors, such as cimetidine and a cationic uremic solutes mixture. The ASP(+) uptake by the living upscaled membrane was decreased by 60% in the presence of either inhibitor, proving the active function of OCT2. In conclusion, this study presents a successful upscaling of a living membrane with active organic cation transport as a support for BAK device. (C) 2016 Elsevier B.V. All rights reserved.