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首页> 外文期刊>Biomaterials >High-throughput screening of microscale pitted substrate topographies for enhanced nonviral transfection efficiency in primary human fibroblasts.
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High-throughput screening of microscale pitted substrate topographies for enhanced nonviral transfection efficiency in primary human fibroblasts.

机译：高通量筛选微凹坑底物形貌，以增强人类原代成纤维细胞的非病毒转染效率。

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摘要

Optimization of nonviral gene delivery typically focuses on the design of particulate carriers that are endowed with desirable membrane targeting, internalization, and endosomal escape properties. Topographical control of cell transfectability, however, remains a largely unexplored parameter. Emerging literature has highlighted the influence of cell-topography interactions on modulation of many cell phenotypes, including protein expression and cytoskeletal behaviors implicated in endocytosis. Using high-throughput screening of primary human dermal fibroblasts cultured on a combinatorial library of microscale topographies, we have demonstrated an improvement in nonviral transfection efficiency for cells cultured on dense micropit patterns compared to smooth substrates, as verified with flow cytometry. A 25% increase in GFP(+) cells was observed independent of proliferation rate, accompanied by SEM and confocal microscopy characterization to help explain the phenomenon qualitatively. This finding encourages researchers to investigate substrate topography as a new design consideration for the optimization of nonviral transfection systems.
机译：非病毒基因传递的优化通常集中在颗粒载体的设计上，这些载体具有理想的膜靶向，内在化和内体逃逸特性。然而，细胞转染性的地形控制仍然是很大程度上未探索的参数。新兴文献强调了细胞-地形学相互作用对许多细胞表型调节的影响，包括与内吞作用有关的蛋白质表达和细胞骨架行为。使用高通量筛选在微尺度地形图组合库中培养的人类原始皮肤成纤维细胞，我们证明了与光滑底物相比，在密集微坑模式下培养的细胞的非病毒转染效率有所提高，这已通过流式细胞仪进行了验证。观察到GFP（+）细胞增加25％，与增殖速率无关，并伴有SEM和共聚焦显微镜表征，以帮助定性解释该现象。这一发现鼓励研究人员研究底物的形貌，将其作为优化非病毒转染系统的新设计考虑。

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