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首页> 外文期刊>Physical chemistry chemical physics: PCCP >Facile preparation of a photoactivatable surface on a 96-well plate: a versatile and multiplex cell migration assay platform
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Facile preparation of a photoactivatable surface on a 96-well plate: a versatile and multiplex cell migration assay platform

机译：在96孔板上轻松制备可光活化表面：多功能的多元细胞迁移测定平台

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Cell migration is an essential cellular activity in various physiological and pathological processes, such as wound healing and cancer metastasis. Therefore, in vitro cell migration assays are important not only for fundamental biological studies but also for evaluating potential drugs that control cell migration activity in medical applications. In this regard, robust control over cell migrating microenvironments is critical for reliable and quantitative analysis as cell migration is highly dependent upon the microenvironments. Here, we developed a facile method for making a commercial glass-bottom 96-well plate photoactivatable for cell adhesion, aiming to develop a versatile and multiplex cell migration assay platform. Cationic poly-D-lysine was adsorbed to the anionic glass surface via electrostatic interactions and, subsequently, functionalized with poly(ethylene glycol) (PEG) bearing a photocleavable reactive group. The initial PEGylated surface is non-cell-adhesive. However, upon near-ultraviolet (UV) irradiation, the photorelease of PEG switches the surface from non-biofouling to cell-adhesive. With this platform, we assayed cell migration in the following procedure: (1) create cell-attaching regions of precise geometries by controlled photo-irradiation, (2) seed cells to allow them to attach selectively to the irradiated regions, (3) expose UV light to the remaining PEGylated regions to extend the cell-adhesive area, (4) analyse cell migration using microscopy. Surface modification of the glass surface was characterized by zeta-potential and contact angle measurements. The PEGylated surface showed cell-resistivity and became cell-adhesive upon releasing PEG by near-UV irradiation. The method was applied for parallelly evaluating the effect of model drugs on the migration of epithelial MDCK cells in the multiplexed platform. The dose-response relationship for cytochalasin D treatment on cell migration behavior was successfully evaluated with high reproducibility. Interestingly, the impact of blebbistatin on cell migration was dependent upon the widths of the migrating regions, resulting in both cases of migration acceleration and deceleration. These results clearly demonstrate that the cellular response to certain drugs is highly affected by their migrating geometries. Therefore, the obtained novel photoactivatable 96-well plate serves as a useful high-throughput platform for the identification of drug candidates that have an effect on cell migration behavior.
机译：细胞迁移是各种生理和病理过程（如伤口愈合和癌症转移）中必不可少的细胞活动。因此，体外细胞迁移测定不仅对于基础生物学研究很重要，而且对于评估在医学应用中控制细胞迁移活性的潜在药物也很重要。在这方面，由于细胞迁移高度依赖于微环境，因此对细胞迁移微环境的严格控制对于可靠和定量分析至关重要。在这里，我们开发了一种简便的方法来制造可光活化细胞粘附的商业化玻璃底96孔板，旨在开发一种多功能的多重细胞迁移测定平台。阳离子聚-D-赖氨酸通过静电相互作用被吸附到阴离子玻璃表面，随后被带有光可裂解反应性基团的聚乙二醇（PEG）官能化。最初的PEG化表面是非细胞粘附的。但是，在近紫外线（UV）照射下，PEG的光释放将表面从非生物污染转换为细胞粘附。使用此平台，我们按照以下步骤分析了细胞迁移：（1）通过受控的光辐照创建精确几何形状的细胞附着区，（2）种子细胞使它们选择性地附着于照射区，（3）暴露紫外线照射到其余的聚乙二醇化区域，以扩展细胞粘附区域，（4）使用显微镜分析细胞迁移。通过ζ电势和接触角测量来表征玻璃表面的表面改性。聚乙二醇化的表面表现出细胞电阻性，并且在通过近紫外线照射释放聚乙二醇后变成细胞粘附性。该方法用于平行评估模型药物对多重平台上皮MDCK细胞迁移的影响。以高重现性成功评估了细胞松弛素D处理对细胞迁移行为的剂量反应关系。有趣的是，blebbistatin对细胞迁移的影响取决于迁移区域的宽度，从而导致迁移加速和减速。这些结果清楚地表明，对某些药物的细胞反应受到其迁移几何形状的高度影响。因此，获得的新型可光激活的96孔板可作为有用的高通量平台，用于鉴定对细胞迁移行为有影响的候选药物。

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