College of Engineering, and Biodynamic Optical Imaging Center (BIOPIC), Peking University, Beijing, 100871, China.
Lab Chip. 2012 Sep 7;12(17):3063-72. doi: 10.1039/c2lc40192d. Epub 2012 Jun 12.
Epigenetic regulation plays an important role in cell migration. Although many methods have been developed to measure the motility of mammalian cells, accurate quantitative assessments of the migration speed of individual cells remain a major challenge. It is difficult for conventional scratch assays to differentiate proliferation from migration during the so-called wound-healing processes because of the long experimental time required. In addition, it is also challenging to create identical conditions for evaluating cell migration by conventional methods. We developed a microfluidic device with precisely created blanks allowing for robust and reproducible cell migration inside accurately-controlled microenvironments to study the regulatory effect of the epigenetic regulator histone deacetylase 7 (HDAC7) on cell migration. Through analyzing time-lapse imaging of the cells migrating into individual blank regions, we can measure the migration speed parameter for human primary cells within a few hours, eliminating the confounding effect of cell proliferation. We also developed an automatic image analysis and a numeric model-based data fitting to set up an integrated cell migration analysis system at single-cell resolution. Using this system, we measured the motility of primary human umbilical vein endothelial cells (HUVECs) and the migration speed reduction due to the silencing of HDAC7 and various other genes. We showed that the migration behaviour of these human primary cells are clearly regulated by epigenetic mechanisms, demonstrating the great potential of this accurate and robust assay in the fields of quantitatively migration studies and high-throughput screening.
表观遗传调控在细胞迁移中起着重要作用。尽管已经开发出许多方法来测量哺乳动物细胞的迁移能力，但对单个细胞迁移速度的准确定量评估仍然是一个主要挑战。传统的划痕实验由于需要较长的实验时间，因此很难在所谓的伤口愈合过程中区分增殖和迁移。此外，用传统方法评估细胞迁移也具有挑战性，因为难以创建相同的条件。我们开发了一种微流控装置，该装置具有精确创建的空白区域，可以在精确控制的微环境中进行稳健且可重复的细胞迁移，从而研究表观遗传调节剂组蛋白去乙酰化酶 7 (HDAC7) 对细胞迁移的调节作用。通过分析细胞迁移到单个空白区域的延时成像，我们可以在数小时内测量人原代细胞的迁移速度参数，消除细胞增殖的混杂影响。我们还开发了自动图像分析和基于数值模型的数据拟合，以在单细胞分辨率下建立集成的细胞迁移分析系统。使用该系统，我们测量了原代人脐静脉内皮细胞 (HUVEC) 的迁移能力以及沉默 HDAC7 和其他各种基因导致的迁移速度降低。我们表明，这些人原代细胞的迁移行为明显受到表观遗传机制的调控，证明了这种准确稳健的测定方法在定量迁移研究和高通量筛选领域具有巨大潜力。
