Design of a microfluidic device to quantify dynamic intra-nuclear deformation during cell migration through confining environments.
Design of a microfluidic device to quantify dynamic intra-nuclear deformation during cell migration through confining environments.
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DOI：
10.1039/c5ib00200a
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发表时间：
2015-12
期刊：
影响因子：
--
通讯作者：
Lammerding J
中科院分区：
文献类型：
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作者：
Davidson PM;Sliz J;Isermann P;Denais C;Lammerding J
细胞迁移通过组织和间质空间的能力是发育和组织稳态、免疫细胞移动性以及各种人类疾病中的重要因素。在3-D迁移过程中细胞核及其相关层的变形在癌症转移的背景下引起了越来越多的关注，其基本假设是，由于层蛋白A/C水平降低而导致的较软的细胞核可能有助于肿瘤扩散。然而，目前的方法来研究细胞在限制三维（3-D）环境中的迁移是有限的，其不精确的控制限制，生理相关性，和/或与高分辨率成像技术的兼容性。我们描述了一种聚二甲基硅氧烷（PDMS）微流体装置的设计，该装置由具有精确限定的收缩的通道组成，该收缩模仿生理环境，使活细胞和固定细胞的高分辨率成像成为可能。该装置促进容易的细胞加载和快速但持久（>24小时）的趋化梯度形成，而不需要连续灌注。使用该装置，我们获得了详细的，定量测量的动态核变形的细胞迁移通过紧密的空间，揭示不同阶段的核易位通过收缩，屈曲的核板，和严重的核内应变。此外，我们发现，核纤层蛋白A/C缺陷的细胞表现出增加和更多的塑性核变形相比，野生型细胞，但只有最小的变化，在核体积，这意味着低核纤层蛋白A/C水平通过增加核变形能力，而不是可压缩性，促进迁移通过收缩。我们的迁移设备与高分辨率延时成像的集成提供了一种强大的新方法，用于研究各种生理相关应用中的细胞内力学和动力学，从癌细胞入侵到免疫细胞招募。
The ability of cells to migrate through tissues and interstitial space is an essential factor during development and tissue homeostasis, immune cell mobility, and in various human diseases. Deformation of the nucleus and its associated lamina during 3-D migration is gathering increasing interest in the context of cancer metastasis, with the underlying hypothesis that a softer nucleus, resulting from reduced levels of lamin A/C, may aid tumour spreading. However, current methods to study the migration of cells in confining three dimensional (3-D) environments are limited by their imprecise control over the confinement, physiological relevance, and/or compatibility with high resolution imaging techniques. We describe the design of a polydimethylsiloxane (PDMS) microfluidic device composed of channels with precisely-defined constrictions mimicking physiological environments that enable high resolution imaging of live and fixed cells. The device promotes easy cell loading and rapid, yet long-lasting (>24 hours) chemotactic gradient formation without the need for continuous perfusion. Using this device, we obtained detailed, quantitative measurements of dynamic nuclear deformation as cells migrate through tight spaces, revealing distinct phases of nuclear translocation through the constriction, buckling of the nuclear lamina, and severe intranuclear strain. Furthermore, we found that lamin A/C-deficient cells exhibited increased and more plastic nuclear deformations compared to wild-type cells but only minimal changes in nuclear volume, implying that low lamin A/C levels facilitate migration through constrictions by increasing nuclear deformability rather than compressibility. The integration of our migration devices with high resolution time-lapse imaging provides a powerful new approach to study intracellular mechanics and dynamics in a variety of physiologically-relevant applications, ranging from cancer cell invasion to immune cell recruitment.