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Curvature facilitates podocyte culture in a biomimetic platform†
Lab on a Chip ( IF 6.1 ) Pub Date : 2018-09-11 00:00:00 , DOI: 10.1039/c8lc00495a
Anastasia Korolj 1, 2, 2, 3, 4 , Carol Laschinger 2, 3, 4 , Chris James 5, 6, 7, 8 , Erding Hu 5, 6, 7, 8 , Claire Velikonja 1, 2, 3 , Nathaniel Smith 1, 2, 3 , Irene Gu 1, 2, 3 , Samad Ahadian 2, 3, 4 , Robert Willette 5, 6, 7, 8 , Milica Radisic 1, 2, 2, 3, 4 , Boyang Zhang 2, 3, 4
Affiliation  

Most kidney diseases begin with abnormalities in glomerular podocytes, motivating the need for podocyte models to study pathophysiological mechanisms and new treatment options. However, podocytes cultured in vitro face a limited ability to maintain appreciable extents of differentiation hallmarks, raising concerns over the relevance of study results. Many key properties such as nephrin expression and morphology reach plateaus that are far from the in vivo levels. Here, we demonstrate that a biomimetic topography, consisting of microhemispheres arrayed over the cell culture substrate, promotes podocyte differentiation in vitro. We define new methods for fabricating microscale curvature on various substrates, including a thin porous membrane. By growing podocytes on our topographic substrates, we found that these biophysical cues augmented nephrin gene expression, supported full-size nephrin protein expression, encouraged structural arrangement of F-actin and nephrin within the cell, and promoted process formation and even interdigitation compared to the flat substrates. Furthermore, the topography facilitated nephrin localization on curved structures while nuclei lay in the valleys between them. The improved differentiation was also evidenced by tracking barrier function to albumin over time using our custom topomembranes. Overall, our work presents accessible methods for incorporating microcurvature on various common substrates, and demonstrates the importance of biophysical stimulation in supporting higher-fidelity podocyte cultivation in vitro.

中文翻译:

弯曲有助于在仿生平台中培养足细胞

大多数肾脏疾病始于肾小球足细胞异常,促使人们需要使用足细胞模型来研究病理生理机制和新的治疗选择。然而,体外培养的足细胞维持明显程度的分化标志方面面临有限的能力,这引起了对研究结果相关性的担忧。肾素的表达和形态等许多关键特性达到了离体内水平很远的平稳期。在这里,我们证明了仿生地形,由排列在细胞培养底物上的微半球组成,可促进体外足细胞分化。我们定义了在包括薄多孔膜在内的各种基底上制造微尺度曲率的新方法。通过在我们的地形基底上生长足细胞,我们发现这些生物物理线索增强了nephrin基因的表达,支持完整大小的nephrin蛋白表达,促进了F-actin和nephrin在细胞内的结构排列,并促进了过程形成,甚至相互交叉。平面基板。此外,地形有利于肾素在弯曲结构上的定位,而核位于它们之间的谷中。通过使用我们定制的拓扑膜随时间追踪白蛋白的屏障功能,也证明了分化的改善。总体而言,我们的工作提出了将微曲率结合到各种常见基质上的简便方法,并证明了生物物理刺激在支持体外高保真足细胞培养的重要性。
更新日期:2018-09-11
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