Tan John L, Tien Joe, Pirone Dana M, Gray Darren S, Bhadriraju Kiran, Chen Christopher S
Department of Biomedical Engineering, The Johns Hopkins University, Baltimore, MD 21205, USA.
Proc Natl Acad Sci U S A. 2003 Feb 18;100(4):1484-9. doi: 10.1073/pnas.0235407100. Epub 2003 Jan 27.
We describe an approach to manipulate and measure mechanical interactions between cells and their underlying substrates by using microfabricated arrays of elastomeric, microneedle-like posts. By controlling the geometry of the posts, we varied the compliance of the substrate while holding other surface properties constant. Cells attached to, spread across, and deflected multiple posts. The deflections of the posts occurred independently of neighboring posts and, therefore, directly reported the subcellular distribution of traction forces. We report two classes of force-supporting adhesions that exhibit distinct force-size relationships. Force increased with size of adhesions for adhesions larger than 1 microm(2), whereas no such correlation existed for smaller adhesions. By controlling cell adhesion on these micromechanical sensors, we showed that cell morphology regulates the magnitude of traction force generated by cells. Cells that were prevented from spreading and flattening against the substrate did not contract in response to stimulation by serum or lysophosphatidic acid, whereas spread cells did. Contractility in the unspread cells was rescued by expression of constitutively active RhoA. Together, these findings demonstrate a coordination of biochemical and mechanical signals to regulate cell adhesion and mechanics, and they introduce the use of arrays of mechanically isolated sensors to manipulate and measure the mechanical interactions of cells.
我们描述了一种通过使用微制造的弹性体微针状柱阵列来操纵和测量细胞与其下层基质之间机械相互作用的方法。通过控制柱的几何形状,我们在保持其他表面特性不变的情况下改变了基质的顺应性。细胞附着在多个柱上,铺展并使柱发生偏转。柱的偏转独立于相邻柱发生,因此直接反映了牵引力的亚细胞分布。我们报告了两类具有不同力-大小关系的力支撑粘附。对于大于1平方微米的粘附,力随粘附大小增加,而对于较小的粘附则不存在这种相关性。通过控制细胞在这些微机械传感器上的粘附,我们表明细胞形态调节细胞产生的牵引力大小。被阻止在基质上铺展和扁平化的细胞在受到血清或溶血磷脂酸刺激时不会收缩,而铺展的细胞会收缩。通过组成型激活的RhoA的表达挽救了未铺展细胞的收缩性。总之,这些发现证明了生化和机械信号的协调以调节细胞粘附和力学,并且引入了使用机械隔离传感器阵列来操纵和测量细胞的机械相互作用。