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细胞旋转培养舱是一种简单且经济的悬浮培养装置,可向肾近端小管细胞传递流体切应力。

Cell spinpods are a simple inexpensive suspension culture device to deliver fluid shear stress to renal proximal tubular cells.

机构信息

Cell Spinpod LLC, Chapel Hill, NC, 27516, USA.

Nephrology Division, Department of Internal Medicine, Duke University School of Medicine, Durham, NC, 27705, USA.

出版信息

Sci Rep. 2021 Oct 29;11(1):21296. doi: 10.1038/s41598-021-00304-8.

Abstract

Rotating forms of suspension culture allow cells to aggregate into spheroids, prevent the de-differentiating influence of 2D culture, and, perhaps most importantly of all, provide physiologically relevant, in vivo levels of shear stress. Rotating suspension culture technology has not been widely implemented, in large part because the vessels are prohibitively expensive, labor-intensive to use, and are difficult to scale for industrial applications. Our solution addresses each of these challenges in a new vessel called a cell spinpod. These small 3.5 mL capacity vessels are constructed from injection-molded thermoplastic polymer components. They contain self-sealing axial silicone rubber ports, and fluoropolymer, breathable membranes. Here we report the two-fluid modeling of the flow and stresses in cell spinpods. Cell spinpods were used to demonstrate the effect of fluid shear stress on renal cell gene expression and cellular functions, particularly membrane and xenobiotic transporters, mitochondrial function, and myeloma light chain, cisplatin and doxorubicin, toxicity. During exposure to myeloma immunoglobulin light chains, rotation increased release of clinically validated nephrotoxicity cytokine markers in a toxin-specific pattern. Addition of cisplatin or doxorubicin nephrotoxins reversed the enhanced glucose and albumin uptake induced by fluid shear stress in rotating cell spinpod cultures. Cell spinpods are a simple, inexpensive, easily automated culture device that enhances cellular functions for in vitro studies of nephrotoxicity.

摘要

旋转悬浮培养允许细胞聚集形成球体,防止二维培养的去分化影响,也许最重要的是,提供生理相关的、体内水平的切变应力。旋转悬浮培养技术尚未得到广泛应用,很大程度上是因为该培养器皿非常昂贵,使用起来劳动强度大,并且难以扩大规模以应用于工业。我们的解决方案在一种名为细胞旋转培养小室的新型器皿中解决了这些挑战。这些小容量为 3.5 毫升的培养器皿由注塑热塑性聚合物组件构成。它们包含自密封轴向硅橡胶端口和氟聚合物透气膜。在这里,我们报告了细胞旋转培养小室中的流场和应力的双流体模拟。细胞旋转培养小室被用于展示流体切变应力对肾细胞基因表达和细胞功能的影响,特别是对膜和外源性化合物转运蛋白、线粒体功能以及骨髓瘤轻链、顺铂和阿霉素的影响。在暴露于骨髓瘤免疫球蛋白轻链时,旋转以毒素特异性的模式增加了临床上验证的肾毒性细胞因子标志物的释放。添加顺铂或阿霉素肾毒素会逆转旋转细胞旋转培养小室中流体切变应力诱导的葡萄糖和白蛋白摄取的增强。细胞旋转培养小室是一种简单、廉价、易于自动化的培养装置,可增强体外肾毒性研究中的细胞功能。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c4ee/8556299/12bc803aac48/41598_2021_304_Fig1_HTML.jpg

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