Analysis of CCR7 mediated T cell transfectant migration using a microfluidic gradient generator
Analysis of CCR7 mediated T cell transfectant migration using a microfluidic gradient generator
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2015
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Journal of Immunological Methods ; 419 (2015). - S. 9-17. - ISSN 0022-1759. - eISSN 1872-7905
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T lymphocyte migration is crucial for adaptive immunity. Manipulation of signaling molecules controlling cell migration combined with in-vitro cell migration analysis provides a powerful research approach. Microfluidic devices, which can precisely configure chemoattractant gradients and allow quantitative single cell analysis, have been increasingly applied to cell migration and chemotaxis studies. However, there are a very limited number of published studies involving microfluidic migration analysis of genetically manipulated immune cells. In this study, we describe a simple microfluidic method for quantitative analysis of T cells expressing transfected chemokine receptors and other cell migration signaling probes. Using this method, we demonstrated chemotaxis of Jurkat transfectants expressing wild-type or C-terminus mutated CCR7 within a gradient of chemokine CCL19, and characterized the difference in transfectant migration mediated by wild-type and mutant CCR7. The EGFP-tagged CCR7 allows identification of CCR7-expressing transfectants in cell migration analysis and microscopy assessment of CCR7 dynamics. Collectively, our study demonstrated the effective use of the microfluidic method for studying CCR7 mediated T cell transfectant migration. We envision this developed method will provide a useful platform to functionally test various signaling mechanisms at the cell migration level.
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570 Biowissenschaften, Biologie
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WU, Xun, Jiandong WU, Hongzhao LI, Daniel LEGLER, Aaron J. MARSHALL, Francis LIN, 2015. Analysis of CCR7 mediated T cell transfectant migration using a microfluidic gradient generator. In: Journal of Immunological Methods. 419, pp. 9-17. ISSN 0022-1759. eISSN 1872-7905. Available under: doi: 10.1016/j.jim.2015.02.008BibTex
@article{Wu2015Analy-31535, year={2015}, doi={10.1016/j.jim.2015.02.008}, title={Analysis of CCR7 mediated T cell transfectant migration using a microfluidic gradient generator}, volume={419}, issn={0022-1759}, journal={Journal of Immunological Methods}, pages={9--17}, author={Wu, Xun and Wu, Jiandong and Li, Hongzhao and Legler, Daniel and Marshall, Aaron J. and Lin, Francis} }
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