High content imaging in the screening of biomaterial-induced MSC behavior
High content imaging in the screening of biomaterial-induced MSC behavior
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DOI：
10.1016/j.biomaterials.2012.10.035
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发表时间：
2013-02-01
期刊：
影响因子：
14
通讯作者：
de Boer, J.
中科院分区：
文献类型：
--
作者：
Unadkat, H. V.;Groen, N.;de Boer, J.
在与生物材料接触时，细胞和周围组织以由材料的物理化学和机械性质决定的方式响应。传统上，使用报告基因或蛋白质表达的侵入性分析方法来监测细胞反应。这些分析方法涉及评估常用标记物的预定义读数，掩盖细胞中诱导的实际情况。因此，应该设想细胞应答的更广泛的表达谱，这在技术上限制了向更高通量系统的放大。然而，越来越多的人认识到，非侵入性获得的形态学读数与基因表达模式有关。在这里，我们介绍了不同的表面粗糙度的三个PLA表面，通过暴露于不同的持续时间的氧等离子体。将间充质基质细胞的反应与未添加分化剂的光滑未处理的PLA表面进行比较。形态学和全基因组表达谱揭示了潜在的细胞变化，这些变化对于骨、软骨和脂肪形成的常用基因标记物是隐藏的。使用通过高含量成像获得的3个形态测量参数，我们能够建立分类器并区分氧等离子体诱导的改性片材和未改性的PLA片材，其中评估经典候选物错过了这种效果。这种方法显示了在高通量系统中使用非侵入性形态测量数据来筛选生物材料表面的可行性，该生物材料表面指示潜在的遗传生物材料诱导的变化。(C)2012爱思唯尔有限公司保留所有权利。
Upon contact with a biomaterial, cells and surrounding tissues respond in a manner dictated by the physicochemical and mechanical properties of the material. Traditionally, cellular responses are monitored using invasive analytical methods that report the expression of genes or proteins. These analytical methods involve assessing commonly used markers for a predefined readout, masking the actual situation induced in the cells. Hence, a broader expression profile of the cellular response should be envisioned, which technically limits up scaling to higher throughput systems. However, it is increasingly recognized that morphometric readouts, obtained non-invasively, are related to gene expression patterns. Here, we introduced distinct surface roughness to three PLA surfaces, by exposure to oxygen plasma of different duration times. The response of mesenchymal stromal cells was compared to smooth untreated PLA surfaces without the addition of differentiation agents. Morphological and genome wide expression profiles revealed underlying cellular changes which was hidden for the commonly used gene markers for osteo-, chondro- and adipogenesis. Using 3 morphometric parameters, obtained by high content imaging, we were able to build a classifier and discriminate between oxygen plasma-induced modified sheets and non-modified PLA sheets where evaluating classical candidates missed this effect. This approach shows the feasibility to use noninvasive morphometric data in high-throughput systems to screen biomaterial surfaces indicating the underlying genetic biomaterial-induced changes. (C) 2012 Elsevier Ltd. All rights reserved.