A silicone-based stretchable micropost array membrane for monitoring live-cell subcellular cytoskeletal response.
A silicone-based stretchable micropost array membrane for monitoring live-cell subcellular cytoskeletal response.
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DOI：
10.1039/c2lc20896b
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发表时间：
2012-02-21
期刊：
影响因子：
6.1
通讯作者：
Fu J
中科院分区：
文献类型：
--
作者：
Mann JM;Lam RH;Weng S;Sun Y;Fu J
外力越来越多地被认为是细胞结构和功能的主要调节因子，但细胞感知力并将其转化为细胞内生化信号和行为反应（“机械转导”）的潜在机制在很大程度上尚未确定。为了帮助机械转导的机制研究，在此我们设计了一种细胞拉伸装置，其允许对机械刺激和细胞生物力学响应进行定量控制和实时测量。我们的策略涉及集成到可拉伸弹性膜上的硅橡胶弹性体微柱的微制造阵列。通过使用计算机控制的真空，该微柱阵列膜（mPAM）被激活以向附着在微柱上的粘附细胞施加等双轴细胞拉伸力。使用mPAM，我们研究了血管平滑肌细胞（VSMCs）的收缩力的活细胞亚细胞动态响应持续静态等双轴细胞拉伸。我们的数据表明，在响应持续的细胞拉伸，VSMC调节其细胞骨架（CSK）收缩性在一个双相的方式：他们首先急剧增强其收缩，以抵抗快速细胞变形（“硬化”）之前，他们允许缓慢的适应性非弹性CSK重组释放其收缩性（“软化”）。整个单个VSMC的收缩反应是空间不均匀的和力依赖性的。我们的mPAM装置和活细胞亚细胞收缩测量将有助于阐明VSMC中的机械转导系统，从而有助于我们理解压力诱导的血管疾病过程。
External forces are increasingly recognized as major regulators of cellular structure and function, yet the underlying mechanism by which cells sense forces and transduce them into intracellular biochemical signals and behavioral responses (‘mechanotransduction’) is largely undetermined. To aid in the mechanistic study of mechanotransduction, herein we devised a cell stretch device that allowed for quantitative control and real-time measurements of mechanical stimuli and cellular biomechanical responses. Our strategy involved a microfabricated array of silicone elastomeric microposts integrated onto a stretchable elastomeric membrane. By using a computer-controlled vacuum, this micropost array membrane (mPAM) was activated to apply equibiaxial cell stretching forces to adherent cells attached on the microposts. Using the mPAM, we studied live-cell subcellular dynamic responses of contractile forces of vascular smooth muscle cells (VSMCs) to sustained static equibiaxial cell stretches. Our data showed that in response to sustained cell stretches, VSMCs regulated their cytoskeletal (CSK) contractility in a biphasic manner: they first acutely enhanced their contraction to resist rapid cell deformation (‘stiffening’) before they allowed slow adaptive inelastic CSK reorganization to release their contractility (‘softening’). The contractile response across entire single VSMCs was spatially inhomogeneous and force-dependent. Our mPAM device and live-cell subcellular contractile measurements will help elucidate the mechanotransductive system in VSMCs and thus contribute to our understanding of pressure-induced vascular disease processes.
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影响因子：
64.5
作者：
Choquet, D;Felsenfeld, DP;Sheetz, MP
通讯作者：
Sheetz, MP
影响因子：
9.7
作者：
Ahmed, W. W.;Kural, M. H.;Saif, T. A.
通讯作者：
Saif, T. A.
影响因子：
3.4
作者：
Gavara, Nuria;Roca-Cusachs, Pere;Navajas, Daniel
通讯作者：
Navajas, Daniel
DOI：
10.1073/pnas.0408482102
发表时间：
2005-02-15
影响因子：
11.1
作者：
du Roure, O;Saez, A;Ladoux, B
通讯作者：
Ladoux, B
影响因子：
3.4
作者：
Bausch, AR;Möller, W;Sackmann, E
通讯作者：
Sackmann, E