Britland S, Morgan H, Wojiak-Stodart B, Riehle M, Curtis A, Wilkinson C
Department of Electronics, University of Glasgow, United Kingdom.
Exp Cell Res. 1996 Nov 1;228(2):313-25. doi: 10.1006/excr.1996.0331.
Guided cell movement is a fundamental process in development and regeneration. We have used microengineered culture substrates to study the interaction between model topographic and adhesive guidance cues in steering BHK cell orientation. Grooves 0.1, 0.5, 1.0, 3.0, and 6.0 microm deep together with pitch-matched aminosilane tracks 5, 12, 25, 50, and 100 microm wide were fabricated on fused silica substrates using photolithographic and dry-etching techniques. The cues were presented to the cells individually, simultaneously in parallel and orthogonally opposed. Cells aligned most strongly to 25-microm-wide adhesive tracks and to 5-microm-wide, 6-microm-deep grooves. Stress fibers and vinculin were found to align with the adhesive tracks and to the grooves and ridges. Cell alignment was profoundly enhanced on all surfaces that presented both cues in parallel. Cells were able to switch alignment from ridges to grooves, and vice versa, depending on the location of superimposed adhesive tracks. Cells aligned preferentially to adhesive tracks superimposed orthogonally over grooves of matched pitch, traversing numerous grooves and ridges. The strength of the cues was more closely matched on narrower 3- and 6-microm-deep gratings with cells showing evidence of alignment to both cues. Confocal fluorescence microscopy revealed two groups of mutually opposed f-actin stress fibers within the same cell, one oriented with the topographic cues and the other with the adhesive cues. However, the adhesive response was consistently dominant. We conclude that cells are able to detect and respond to multiple guidance cues simultaneously. The adhesive and topographic guidance cues modeled here were capable of interacting both synergistically and hierarchically to guide cell orientation.
引导细胞运动是发育和再生过程中的一个基本过程。我们使用微工程培养底物来研究模型地形和粘附引导线索在引导BHK细胞取向方面的相互作用。利用光刻和干蚀刻技术，在熔融石英底物上制作了深度为0.1、0.5、1.0、3.0和6.0微米的凹槽，以及宽度为5、12、25、50和100微米且间距匹配的氨基硅烷轨道。这些线索分别呈现给细胞，同时以平行和正交相对的方式呈现。细胞对25微米宽的粘附轨道和5微米宽、6微米深的凹槽排列最为强烈。发现应力纤维和纽蛋白与粘附轨道以及凹槽和脊排列一致。在所有平行呈现两种线索的表面上，细胞排列都显著增强。细胞能够根据叠加的粘附轨道的位置，从脊切换到凹槽排列，反之亦然。细胞优先与正交叠加在匹配间距凹槽上的粘附轨道排列，穿过许多凹槽和脊。在较窄的3微米和6微米深的光栅上，线索的强度更紧密匹配，细胞显示出对两种线索都有排列的迹象。共聚焦荧光显微镜显示，在同一细胞内有两组相互对立的f-肌动蛋白应力纤维，一组与地形线索取向一致，另一组与粘附线索取向一致。然而，粘附反应始终占主导地位。我们得出结论，细胞能够同时检测并对多种引导线索做出反应。这里模拟的粘附和地形引导线索能够协同和分层相互作用，以引导细胞取向。
