Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Proc Natl Acad Sci U S A. 2011 Jul 5;108(27):11115-20. doi: 10.1073/pnas.1103581108. Epub 2011 Jun 20.
Interstitial flow is the convective transport of fluid through tissue extracellular matrix. This creeping fluid flow has been shown to affect the morphology and migration of cells such as fibroblasts, cancer cells, endothelial cells, and mesenchymal stem cells. A microfluidic cell culture system was designed to apply stable pressure gradients and fluid flow and allow direct visualization of transient responses of cells seeded in a 3D collagen type I scaffold. We used this system to examine the effects of interstitial flow on cancer cell morphology and migration and to extend previous studies showing that interstitial flow increases the metastatic potential of MDA-MB-435S melanoma cells [Shields J, et al. (2007) Cancer Cell 11:526-538]. Using a breast carcinoma line (MDA-MB-231) we also observed cell migration along streamlines in the presence of flow; however, we further demonstrated that the strength of the flow as well as the cell density determined directional bias of migration along the streamline. In particular, we found that cells either at high seeding density or with the CCR-7 receptor inhibited migration against, rather than with the flow. We provide further evidence that CCR7-dependent autologous chemotaxis is the mechanism that leads to migration with the flow, but also demonstrate a competing CCR7-independent mechanism that causes migration against the flow. Data from experiments investigating the effects of cell concentration, interstitial flow rate, receptor activity, and focal adhesion kinase phosphorylation support our hypothesis that the competing stimulus is integrin mediated. This mechanism may play an important role in development of metastatic disease.
间质流是指通过组织细胞外基质的对流式液体传输。这种蠕动式流体流动已被证明会影响成纤维细胞、癌细胞、内皮细胞和间充质干细胞等细胞的形态和迁移。本研究设计了一种微流控细胞培养系统,以施加稳定的压力梯度和流体流动,并允许直接观察在 3D 胶原 I 支架中接种的细胞的瞬态响应。我们使用该系统研究了间质流对癌细胞形态和迁移的影响,并扩展了先前的研究结果,即间质流增加了 MDA-MB-435S 黑色素瘤细胞的转移潜力[Shields J,等人。(2007)Cancer Cell 11:526-538]。我们还使用乳腺癌细胞系(MDA-MB-231)观察到在存在流动的情况下细胞沿流线迁移;然而,我们进一步证明,流动的强度以及细胞密度决定了沿流线迁移的方向偏差。特别是,我们发现,在高接种密度或 CCR-7 受体存在的情况下,细胞会迁移抵抗而不是顺应流动。我们提供了进一步的证据,表明 CCR7 依赖性自体趋化作用是导致与流动相适应的迁移的机制,但也证明了一种竞争的 CCR7 独立机制,导致细胞迁移与流动相背。研究细胞浓度、间质流速、受体活性和粘着斑激酶磷酸化作用的实验数据支持我们的假设,即竞争刺激是由整合素介导的。这种机制可能在转移性疾病的发展中起重要作用。