TopoWellPlate: A Well-Plate-Based Screening Platform to Study Cell-Surface Topography Interactions
TopoWellPlate: A Well-Plate-Based Screening Platform to Study Cell-Surface Topography Interactions
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DOI：
10.1002/adbi.201700002
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发表时间：
2017-04-01
影响因子：
4.1
通讯作者：
de Boer, Jan
中科院分区：
文献类型：
--
作者：
Beijer, Nick R. M.;Vasilevich, Aliaksei S.;de Boer, Jan
生物材料工程领域越来越多地使用高通量方法来研究细胞-材料相互作用。由于大多数材料库都是作为芯片制备的，因此基于免疫荧光的读数用于对单个材料进行独特的成像。本文建议使用一种基于良好的策略来生产材料库，其中每种材料都是物理分离的，因此与标准的生化分析相兼容。在这项工作中，TopoWellPlate，一个新的系统，研究细胞表面形貌相互作用的高通量。从一个更大的地形库中，鉴定了87种独特定义的生物活性表面地形，它们诱导了各种各样的细胞形态。地形增强聚苯乙烯薄膜在多步骤洁净室工艺中制造，并作为TopoWellPlate的基础。薄膜与无底96孔板的热键合产生细胞培养准备，地形增强，96孔板。测量骨髓来源的人间充质干细胞的总体代谢活性，以显示TopoWellPlate作为筛选工具的功能，显示每个细胞的代谢活性差异范围为2.5倍。这种和其他地形设计的topowellplate可用于使用丰富的标准化分子测定来分析细胞，从而揭示生物材料诱导的机械转导机制。
The field of biomaterial engineering is increasingly using high-throughput approaches to investigate cell-material interactions. Because most material libraries are prepared as chips, immunofluorescence-based read-outs are used to uniquely image individual materials. This paper proposes to produce libraries of materials using a well-based strategy in which each material is physically separated, and thus compatible with standard biochemical assays. In this work, the TopoWellPlate, a novel system to study cell-surface topography interaction in high-throughput is presented. From a larger library of topographies, 87 uniquely defined bioactive surface topographies are identified, which induce a wide variety of cellular morphologies. Topographically enhanced polystyrene films are fabricated in a multistep clean room process and served as base for the TopoWellPlate. Thermal bonding of the films to bottomless 96-well plates results in a cell culture ready, topographically enhanced, 96-well plate. The overall metabolic activity of bone marrow-derived human mesenchymal stem cells is measured to show the functionality of the TopoWellPlate as a screening tool, which showed a 2.5-fold difference range in metabolic activity per cell. TopoWellPlates of this and other topographical designs can be used to analyze cells using the wealth of standardized molecular assays available and thus disclose the mechanisms of biomaterials-induced mechanotransduction.