Reprogramming cellular phenotype by soft collagen gels.
Reprogramming cellular phenotype by soft collagen gels.
复制标题
DOI：
10.1039/c4sm01602e
复制
发表时间：
2014-11-28
期刊：
影响因子：
3.4
通讯作者：
Saif MT
中科院分区：
文献类型：
--
作者：
Ali MY;Chuang CY;Saif MT
多种细胞类型响应于基底的机械刚度而表现出表型变化。许多细胞（不包括神经元）显示扩散面积增加，肌动蛋白应力纤维形成和更大的粘着斑复合物作为基板刚度增加，在人口稀少的文化。还已知细胞增殖与这些表型变化/基质硬度变化直接相关。在更硬的基底上增强的铺展和增殖需要核转录调节因子雅普（Yes associated protein）在细胞核中的定位，并且与更大的牵引力产生紧密耦合。在这项研究中，我们表明，不同类型的成纤维细胞可以表现出蔓延形态，明确的肌动蛋白应力纤维，和更大的粘着斑7非常软的胶原蛋白凝胶（模量在数百帕斯卡），就好像他们是在硬玻璃基板（模量在GPa，几个数量级更高）。引人注目的是，我们表明，第一次，软胶原蛋白凝胶上的增强扩散和其他硬基质细胞骨架结构与细胞增殖模式无关，不需要雅普定位在细胞核中。最后，我们研究了人结肠癌（HCT-8）细胞对软胶原凝胶的反应。最近的研究表明，人结肠癌（HCT-8）细胞在具有宽范围刚度（0.5 - 50 kPa）并涂覆有细胞外基质ECM（胶原单体/纤连蛋白）的软聚丙烯酰胺（PA）凝胶上培养2-3天时形成多细胞簇。这些簇在PA凝胶上显示出有限的扩散/润湿，在边缘形成3D结构，并最终显示出显著的解离性转移样表型（MLP），即，仅在PA凝胶上培养一周后，上皮细胞向圆形形态转变，但在胶原单体包被的硬聚苯乙烯/玻璃上没有，在那里它们表现出增强的润湿性并形成汇合的单层。在这里，我们发现HCT-8细胞簇在软胶原凝胶上也表现出增强的扩散/湿润，并最终形成与在刚性玻璃基底上一样的融合单层，并且MLP在软胶原凝胶上被完全抑制。总的来说，这些结果表明，细胞-材料相互作用（在这种情况下，软胶原凝胶）可以诱导细胞表型和细胞骨架组织在一个显着不同的方式相比，一个经典的合成聚丙烯酰胺（PA）水凝胶细胞培养模型，并可能有助于设计新的功能性生物材料。
A variety of cell types exhibit phenotype changes in response to the mechanical stiffness of the substrate. Many cells excluding neurons display increase in spread area, actin stress fiber formation and larger focal adhesion complexes as substrate stiffness increases in sparsely populated culture. Cell proliferation is also known to directly correlate with these phenotype changes/change in substrate stiffness. Augmented spreading and proliferation on stiffer substrates require nuclear transcriptional regulator YAP (Yes associated protein) localization in cell nucleus and is tightly coupled with larger traction force generation. In this study, we show that different types of fibroblasts can exhibit spread morphology, well defined actin stress fibers, and larger focal adhesion seven on very soft collagen gels (modulus in hundreds of Pascals)as if they are on hard glass substrate (modulus in GPa, several orders of magnitude higher). Strikingly, we show, for the first time, that augmented spreading and other hard substrate cytoskeleton architecture on soft collagen gels are not correlated with cell proliferation pattern and do not require YAP localization in cell nucleus. Finally, we examine the response of human colon carcinoma (HCT-8) cells on soft collagen gels. Recent studies show that human colon carcinoma (HCT-8) cells form multi cellular clusters by 2–3 days when cultured on soft polyacrylamide (PA) gels with a wide range of stiffness (0.5 – 50 kPa) and coated with extracellular matrix, ECM (collagen monomer/ fibronectin). These clusters show limited spreading/wetting on PA gels, form 3D structures at the edges, and eventually display a remarkable, dissociative metastasis like phenotype (MLP), i.e., epithelial to rounded morphological transition after a week of culture on PA gels only, but not on collagen monomer coated stiff polystyrene/glass where they exhibit enhanced wetting and form confluent monolayer. Here, we show that HCT-8 cell clusters also show augmented spreading/wetting on soft collagen gels and eventually form confluent monolayer as on rigid glass substrates and MLP is completely inhibited on soft collagen gels. Overall, these results suggest that cell-material interaction (soft collagen gels in this case) can induce cellular phenotype and cytoskeleton organization in a remarkably distinct manner compared to a classical synthetic polyacrylamide (PA) hydrogel cell culture model and may contribute in designing new functional biomaterials.
登录
查看更多内容
影响因子：
3.4
作者：
Chou, Szu-Yuan;Cheng, Chao-Min;LeDuc, Philip R.
通讯作者：
LeDuc, Philip R.
影响因子：
2.1
作者：
Allen, John R.;Ross, Stephen T.;Davidson, Michael W.
通讯作者：
Davidson, Michael W.
影响因子：
3.7
作者：
Baker EL;Srivastava J;Yu D;Bonnecaze RT;Zaman MH
通讯作者：
Zaman MH
影响因子：
2.8
作者：
Ali, M. Yakut;Saif, M. Taher A.
通讯作者：
Saif, M. Taher A.
影响因子：
6.6
作者：
Saunders, Ruth M.;Holt, Mark R.;Critchley, David R.
通讯作者：
Critchley, David R.