Systematic Analysis of In Vitro Cell Rolling Using a Multi-well Plate Microfluidic System |
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Authors: | Oren Levy Priya Anandakumaran Jessica Ngai Rohit Karnik Jeffrey M Karp |
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Institution: | 1.Division of Biomedical Engineering, Department of Medicine, Brigham and Women''s Hospital;2.Center for Regenerative Therapeutics, Brigham and Women''s Hospital;3.Harvard Medical School, Harvard University;4.Harvard Stem Cell Institute, Harvard University;5.Harvard-MIT Division of Health Sciences and Technology;6.Department of Mechanical Engineering, Massachusetts Institute of Technology |
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Abstract: | A major challenge for cell-based therapy is the inability to systemically target a large quantity of viable cells with high efficiency to tissues of interest following intravenous or intraarterial infusion. Consequently, increasing cell homing is currently studied as a strategy to improve cell therapy. Cell rolling on the vascular endothelium is an important step in the process of cell homing and can be probed in-vitro using a parallel plate flow chamber (PPFC). However, this is an extremely tedious, low throughput assay, with poorly controlled flow conditions. Instead, we used a multi-well plate microfluidic system that enables study of cellular rolling properties in a higher throughput under precisely controlled, physiologically relevant shear flow1,2. In this paper, we show how the rolling properties of HL-60 (human promyelocytic leukemia) cells on P- and E-selectin-coated surfaces as well as on cell monolayer-coated surfaces can be readily examined. To better simulate inflammatory conditions, the microfluidic channel surface was coated with endothelial cells (ECs), which were then activated with tumor necrosis factor-α (TNF-α), significantly increasing interactions with HL-60 cells under dynamic conditions. The enhanced throughput and integrated multi-parameter software analysis platform, that permits rapid analysis of parameters such as rolling velocities and rolling path, are important advantages for assessing cell rolling properties in-vitro. Allowing rapid and accurate analysis of engineering approaches designed to impact cell rolling and homing, this platform may help advance exogenous cell-based therapy. |
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Keywords: | Bioengineering Issue 80 Microfluidics Endothelial Cells Leukocyte Rolling HL-60 cells TNF-α P-selectin E-selectin |
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