Multifunctional polymer scaffolds with adjustable pore size and chemoattractant gradients for studying cell matrix invasion
Multifunctional polymer scaffolds with adjustable pore size and chemoattractant gradients for studying cell matrix invasion
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DOI：
10.1016/j.biomaterials.2013.09.095
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发表时间：
2014-01-01
期刊：
影响因子：
14
通讯作者：
Bastmeyer, Martin
中科院分区：
文献类型：
--
作者：
Greiner, Alexandra M.;Jaeckel, Maria;Bastmeyer, Martin
迁移的细胞通常需要使细胞体和细胞核变形以穿过细胞外基质支架中的微米级孔。此外，化学吸引信号通常引导迁移，但在3D基质侵入期间机械约束和信号传导机制之间的精确相互作用尚未完全理解，并且可能在细胞类型之间存在差异。在这里，我们使用直接激光写入在微孔载体膜上制造具有可调节孔径（2-10 μ m）的3D细胞培养支架，用于施加可扩散的化学梯度。小鼠胚胎成纤维细胞侵入10 gm孔支架，即使在没有化学引诱剂，但入侵显着增强敲除核纤层蛋白A/C，一个已知的调节细胞核硬度。因此，核硬度构成了成纤维细胞侵袭基质的主要障碍，但趋化性信号不是必需的。相反，上皮A549细胞即使在核纤层蛋白A/C水平降低时也不进入10 μ m的孔，但在化学引诱物（血清）存在下容易进入孔低至7 μ m的支架。因此，核硬度不是上皮细胞中基质侵入的主要调节因子，而是需要化学吸引信号。因此，具有可调节的孔径和可扩散的化学梯度的微结构支架是在基质侵入期间解剖细胞类型特异性机械和信号方面的有价值的工具。(C)2013爱思唯尔有限公司保留所有权利。
Transmigrating cells often need to deform cell body and nucleus to pass through micrometer-sized pores in extracellular matrix scaffolds. Furthermore, chemoattractive signals typically guide transmigration, but the precise interplay between mechanical constraints and signaling mechanisms during 3D matrix invasion is incompletely understood and may differ between cell types. Here, we used Direct Laser Writing to fabricate 3D cell culture scaffolds with adjustable pore sizes (2-10 mu m) on a microporous carrier membrane for applying diffusible chemical gradients. Mouse embryonic fibroblasts invade 10 gm pore scaffolds even in absence of chemoattractant, but invasion is significantly enhanced by knockout of lamin A/C, a known regulator of cell nucleus stiffness. Nuclear stiffness thus constitutes a major obstacle to matrix invasion for fibroblasts, but chemotaxis signals are not essential. In contrast, epithelial A549 cells do not enter 10 mu m pores even when lamin A/C levels are reduced, but readily enter scaffolds with pores down to 7 mu m in presence of chemoattractant (serum). Nuclear stiffness is therefore not a prime regulator of matrix invasion in epithelial cells, which instead require chemoattractive signals. Micro-structured scaffolds with adjustable pore size and diffusible chemical gradients are thus a valuable tool to dissect cell-type specific mechanical and signaling aspects during matrix invasion. (C) 2013 Elsevier Ltd. All rights reserved.