Traction Force Screening Enabled by Compliant PDMS Elastomers.
Traction Force Screening Enabled by Compliant PDMS Elastomers.
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由合规 PDMS 弹性体实现牵引力筛选。
DOI：
10.1016/j.bpj.2018.02.045
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发表时间：
2018
影响因子：
3.4
通讯作者：
Ehrlicher,AllenJ
中科院分区：
文献类型：
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作者：
Yoshie,Haruka;Koushki,Newsha;Kaviani,Rosa;Tabatabaei,Mohammad;Rajendran,Kavitha;Dang,Quynh;Husain,Amjad;Yao,Sean;Li,Chuck;Sullivan,JohnK;Saint-Geniez,Magali;Krishnan,Ramaswamy;Ehrlicher,AllenJ
肌动球蛋白收缩性是细胞生物学许多方面的基本要素，并表现为细胞对其周围环境施加的牵引力。这些力量的核心作用使它们成为各种疾病的新的主要治疗靶点。这需要精确和更高容量的牵引力测量;然而，现有的方法在很大程度上是低吞吐量，限制了它们在更广泛的应用中的效用。为了满足这一需求，我们采用傅立叶变换牵引力显微镜在一个平行的96孔格式，我们称之为收缩力筛选。重要的是，而不是经常使用的水凝胶聚丙烯酰胺，我们制造这些板使用聚二甲基硅氧烷橡胶。这种方法的关键是所使用的聚二甲基硅氧烷非常柔顺，下限杨氏模量为10.4 kPa。我们将这些整体式基质在空间上细分为生物化学上独立的威尔斯，从而创建用于牵引力筛选的统一多孔平台。我们通过量化人气道平滑肌细胞和视网膜色素上皮细胞的化合物和剂量依赖性收缩反应来证明该平台的实用性和多功能性。通过直接量化治疗意图的终点，气道平滑肌收缩力，该方法填补了目前用于支气管扩张药物发现的筛选方法中的重要方法学空白，并且更一般地，在测量广泛的细胞类型和病理的收缩反应中。
Actomyosin contractility is an essential element of many aspects of cellular biology and manifests as traction forces that cells exert on their surroundings. The central role of these forces makes them a novel principal therapeutic target in diverse diseases. This requires accurate and higher-capacity measurements of traction forces; however, existing methods are largely low throughput, limiting their utility in broader applications. To address this need, we employ Fourier-transform traction force microscopy in a parallelized 96-well format, which we refer to as contractile force screening. Critically, rather than the frequently employed hydrogel polyacrylamide, we fabricate these plates using polydimethylsiloxane rubber. Key to this approach is that the polydimethylsiloxane used is very compliant, with a lower-bound Young's modulus of ∼0.4 kPa. We subdivide these monolithic substrates spatially into biochemically independent wells, creating a uniform multiwell platform for traction force screening. We demonstrate the utility and versatility of this platform by quantifying the compound and dose-dependent contractility responses of human airway smooth muscle cells and retinal pigment epithelial cells. By directly quantifying the endpoint of therapeutic intent, airway-smooth-muscle contractile force, this approach fills an important methodological void in current screening approaches for bronchodilator drug discovery, and, more generally, in measuring contractile response for a broad range of cell types and pathologies.