Patterning cells in highly deformable micro structures: Effect of plastic deformation of substrate on cellular phenotype and gene expression
Patterning cells in highly deformable micro structures: Effect of plastic deformation of substrate on cellular phenotype and gene expression
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DOI：
10.1016/j.biomaterials.2005.08.018
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发表时间：
2006-03-01
期刊：
影响因子：
14
通讯作者：
Chilkoti, A
中科院分区：
文献类型：
--
作者：
Hyun, JH;Chen, J;Chilkoti, A
我们描述了可变形的微结构的制造低压软微压花（亩SEMB），提供了在体外实验的“试验台”，以调查在一个高度受控的环境中的细胞行为的机械和化学刺激的相互作用。软微压印利用石蜡膜的柔软性和可塑性，通过将硅母模或弹性体聚（二甲基硅氧烷）印模压入石蜡膜中来制造微结构。我们证明，抗蛋白质梳形聚合物可以被印刷到凸起的特征的压花的微结构，赋予蛋白质，因此细胞阻力的微结构的那些区域。这两个特点，我们的制造方法-微压花，然后空间选择性转移的无污染的聚合物-形成我们的战略的核心，以图案化的石蜡膜微结构内的细胞，使细胞被限制在底部的微结构。细胞培养实验表明，NIH 3 T3成纤维细胞的优先细胞附着的纤连蛋白（FN）的免疫荧光显微镜的微图案。肌动蛋白细胞骨架重新排列沿着轴施加机械应力，拉伸细胞显示改变的基因表达的细胞骨架和基质蛋白在响应机械变形。使用parafilm作为基板和mu SEMB作为制造方法提供了一个简单的和广泛访问的方法来研究细胞行为tinder良好定义的条件下的塑性变形和表面配体密度。(c)2005爱思唯尔有限公司保留所有权利。
We describe the fabrication of deformable microstructures by low-pressure-soft-microembossing (mu SEmb) that provides in vitro experimental "test-beds" to investigate the interplay of mechanical and chemical stimuli on cell behavior in a highly controlled environment. Soft microembossing exploits the softness and plasticity of parafilm to fabricate microstructures by pressing a silicon master or an elastomeric poly(dimethylsiloxane) stamp into the parafilm. We demonstrate that a protein-resistant comb polymer can be printed into the raised features of the embossed micro structures, which imparts protein, and hence cell resistance to those regions of the microstructures. These two features of our fabrication methodology-microembossing followed by spatially selective transfer of a nonfouling polymer-forms the core of our strategy to pattern cells within the parafilm microstructures, such that the cells are confined within bottoms of the microstructures. Cell culture experiments demonstrated the preferential cell attachment of NIH 3T3 fibroblasts to the fibronectin (FN) micropatterns by immunofluorescence microscopy. The actin cytoskeleton realigned along the axis of applied mechanical stress, and stretched cells showed altered gene expression of cytoskeletal and matrix proteins in response to mechanical deformation. The use of parafilm as a substrate and mu SEmb as a fabrication method provides a simple and widely accessible methodology to investigate cellular behavior tinder well-defined conditions of plastic deformation and surface ligand density. (c) 2005 Elsevier Ltd. All rights reserved.