Micropatterning tractional forces in living cells
Micropatterning tractional forces in living cells
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DOI：
10.1002/cm.10037
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发表时间：
2002-06-01
影响因子：
--
通讯作者：
Ingber, DE
中科院分区：
文献类型：
--
作者：
Wang, N;Ostuni, E;Ingber, DE
在这里，我们描述了一种量化细胞牵引力的方法，这些细胞在物理上被限制在含有荧光微珠（直径为 0.2 微米）的柔性聚丙烯酰胺凝胶表面上具有限定形状和尺寸的微米级粘合岛内。将平滑肌细胞铺在方形（50 x 50 mm）或圆形（直径 25 或 50 mm）粘合岛上，这些粘合岛是通过在弹性体膜上的微工程孔涂上胶原蛋白涂层而形成的，随后将其去除。贴壁细胞扩散至岛的大小和形状，并通过绘制凝胶内荧光微珠的位移场来定量细胞牵引力。圆形岛上的细胞没有表现出任何优先的施力方向，但它们在形成突起的部位发挥了最强的牵引力。当细胞被限制在正方形内时，无论是否存在收缩激动剂、组胺，角落处的牵引力最高，并且在这些区域中也观察到细胞突起。对不同岛上培养的细胞施加的平均牵引力的定量表明，随着细胞扩散的促进，细胞张力增加。这些结果为过去的研究提供了机械基础，这些研究证明了各种贴壁依赖性细胞内的扩散和生长之间存在类似的相关性。这种用于分析单个细胞下方机械力空间分布的新方法可以为细胞调节的生物物理基础提供更多见解。 (C) 2002 Wiley-Liss, Inc.
Here we describe a method for quantifying traction in cells that are physically constrained within micron-sized adhesive islands of defined shape and size on the surface of flexible polyacrylamide gels that contain fluorescent microbeads (0.2-mum diameter). Smooth muscle cells were plated onto square (50 x 50 mum) or circular (25- or 50-mum diameter) adhesive islands that were created on the surface of the gels by applying a collagen coating through microengineered holes in an elastomeric membrane that was later removed. Adherent cells spread to take on the size and shape of the islands and cell tractions were quantitated by mapping displacement fields of the fluorescent microbeads within the gel. Cells on round islands did not exhibit any preferential direction of force application, but they exerted their strongest traction at sites where they formed protrusions. When cells were confined to squares, traction was highest in the corners both in the absence and presence of the contractile agonist, histamine, and cell protrusions were also observed in these regions. Quantitation of the mean traction exerted by cells cultured on the different islands revealed that cell tension increased as cell spreading was promoted. These results provide a mechanical basis for past studies that demonstrated a similar correlation between spreading and growth within various anchorage-dependent cells. This new approach for analyzing the spatial distribution of mechanical forces beneath individual cells that are experimentally constrained to defined sizes and shapes may provide additional insight into the biophysical basis of cell regulation. (C) 2002 Wiley-Liss, Inc.