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首页> 外文期刊>Cell motility and the cytoskeleton >Direct, dynamic assessment of cell-matrix interactions inside fibrillar collagen lattices.
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Direct, dynamic assessment of cell-matrix interactions inside fibrillar collagen lattices.

机译：直接，动态评估纤维状胶原蛋白晶格内部的细胞-基质相互作用。

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Cell mechanical behavior has traditionally been studied using 2-D planar elastic substrates. The goal of this study was to directly assess cell-matrix mechanical interactions inside more physiologic 3-D collagen matrices. Rabbit corneal fibroblasts transfected to express GFP-zyxin were plated at low density inside 100 micro m-thick type I collagen matrices. 3-D datasets of isolated cells were acquired at 1-3-min intervals for up to 5 h using fluorescent and Nomarski DIC imaging. Unlike cells on 2-D substrates, cells inside the collagen matrices had a bipolar morphology with thin pseudopodial processes, and without lamellipodia. The organization of the collagen fibrils surrounding each cell was clearly visualized using DIC. Using time-lapse color overlays of GFP and DIC images, displacement and/or realignment of collagen fibrils by focal adhesions could be directly visualized. During pseudopodial extension, new focal adhesions often formed in a line along collagen fibrils in front of the cell, while existing adhesions moved backward. This process generated tractional forces as indicated by the pulling in of collagen fibrils in front of the cell. Meanwhile, adhesions on both the dorsal and ventral surface of the cell body generally moved forward, resulting in contractile shortening along the pseudopodia and localized extracellular matrix (ECM) compression. Cytochalasin D induced rapid disassembly of focal adhesions, cell elongation, and ECM relaxation. This experimental model allows direct, dynamic assessment of cell-matrix interactions inside a 3-D fibrillar ECM. The data suggest that adhesions organize along actin-based contractile elements that are much less complex than the network of actin filaments that mechanically links lamellar adhesions on 2-D substrates.
机译：传统上已经使用二维平面弹性基板研究了电池的机械行为。这项研究的目的是直接评估更多生理3D胶原蛋白基质内的细胞基质机械相互作用。将转染以表达GFP-zyxin的兔角膜成纤维细胞以低密度接种在100微米m厚的I型胶原蛋白基质中。使用荧光和Nomarski DIC成像，以1-3分钟的间隔获取长达5小时的分离细胞的3-D数据集。与2-D底物上的细胞不同，胶原蛋白基质内的细胞具有双极形态，具有较薄的假足突，并且没有片状脂膜。使用DIC可以清楚地看到每个细胞周围的胶原蛋白原纤维的组织。使用GFP和DIC图像的延时彩色叠加，可以直接观察到粘着斑对胶原纤维的位移和/或重新排列。在假足伸展过程中，新的粘着斑通常沿着细胞前的胶原蛋白原纤维成一条线，而现有的粘着斑则向后移动。这个过程产生了牵引力，如胶原蛋白原纤维在细胞前部的拉入所表明的。同时，在细胞体的背侧和腹侧表面上的粘附通常向前移动，导致沿伪足的收缩缩短和局部细胞外基质（ECM）压缩。细胞松弛素D引起粘着斑快速分解，细胞伸长和ECM松弛。该实验模型可以直接，动态地评估3-D纤维状ECM内部的细胞-基质相互作用。数据表明，粘附力沿着肌动蛋白基收缩元件组织，而肌动蛋白细丝的网络比将肌动蛋白丝机械连接到二维基底上的层状粘附力复杂得多。

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