Live cell imaging analysis of the epigenetic regulation of the human endothelial cell migration at single-cell resolution.

Chunhong Zheng, Zhilong Yu, Ying Zhou, Louis Tao, Yuhong Pang, Tao Chen, Xiannian Zhang, Haiwei Qiu, Hongwei Zhou, Zitian Chen, Yanyi Huang

College of Engineering, and Biodynamic Optical Imaging Center (BIOPIC), Peking University, Beijing, 100871, China.

Lab on a chip 2012 Sep 7

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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.

Citation

Chunhong Zheng, Zhilong Yu, Ying Zhou, Louis Tao, Yuhong Pang, Tao Chen, Xiannian Zhang, Haiwei Qiu, Hongwei Zhou, Zitian Chen, Yanyi Huang. Live cell imaging analysis of the epigenetic regulation of the human endothelial cell migration at single-cell resolution. Lab on a chip. 2012 Sep 7;12(17):3063-72