Department of Engineering Science, National Cheng Kung University, 1 University Road, Tainan, 701, Taiwan, ROC.
Lab Chip. 2011 Aug 7;11(15):2583-90. doi: 10.1039/c1lc20113a. Epub 2011 Jun 21.
We have successfully developed a microsystem to co-cultivate two types of cells with a minimum defined gap of 50 μm, and to quantitatively study the impact of fluid shear stress on the mutual influence of cell migration velocity and distance. We used the hydrostatic pressure to seed two different cells, endothelial cells (ECs) and smooth muscle cells (SMCs), on opposite sides of various gap sizes (500 μm, 200 μm, 100 μm, and 50 μm). After cultivating the cells for 12 h and peeling the co-culture microchip from the culture dish, we studied the impacts of gap size on the migration of either cell type in the absence or presence of fluid shear stress (7 dyne cm(-2) and 12 dyne cm(-2)) influence. We found that both gap size and shear stress have profound influence on cell migration. Smaller gap sizes (100 μm and 50 μm) significantly enhanced cell migration, suggesting a requirement of an effective concentration of released factor(s) by either cell type in the gap region. Flow-induced shear stress delayed the migration onset of either cell type in a dose-dependent manner regardless of the gap size. Moreover, shear stress-induced decrease of cell migration becomes evident when the gap size was 500 μm. We have developed a co-culture microsystem for two kinds of cells and overcome the conventional difficulties in observation and mixed culture, and it would have more application for bio-manipulation and tissue repair engineering.
我们成功开发了一种微系统，可以共培养两种细胞，两种细胞之间的最小定义间隔为 50μm，并定量研究流体切应力对细胞迁移速度和距离相互影响的影响。我们使用静水压力将两种不同的细胞，内皮细胞（ECs）和平滑肌细胞（SMCs），种植在不同间隔大小（500μm、200μm、100μm 和 50μm）的相对侧。在培养细胞 12 小时并从培养皿中剥离共培养微芯片后，我们研究了在没有或存在流体切应力（7 达因/厘米(-2)和 12 达因/厘米(-2))的情况下，间隔大小对任一种细胞类型迁移的影响。我们发现，间隔大小和切应力都对细胞迁移有深远的影响。较小的间隔大小（100μm 和 50μm）显著增强了细胞迁移，表明在间隔区域中任一种细胞类型都需要释放有效浓度的因子。无论间隔大小如何，流动引起的切应力都以剂量依赖的方式延迟了任一种细胞类型的迁移起始。此外，当间隔大小为 500μm 时，剪切应力诱导的细胞迁移减少变得明显。我们开发了一种用于两种细胞的共培养微系统，克服了观察和混合培养的传统困难，它将在生物操作和组织修复工程中有更多的应用。
