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Characterization and Application of a Long Term Noncytotoxic Live Cell Labeling Probe

Abstract

Fluorescent molecular probes for cell labeling are useful in visualizing and quantifying cells. Many commercially available probes are cytotoxic and not ideal for multi-day live-cell tracking. In order to assess their effects on cell proliferation, the number of cells exposed to a probe was automatically counted every day for five days using an image analysis software called CellProfiler. MitoTracker Red at 62.5 nM showed the least interference with cell growth and produced desirable signals and hence was identified as the most suitable live-cell staining agent for long-term tracking. MitoTracker Red was subsequently used to quantify the effects of honokiol, an extract from a plant, on vascular cell proliferation. Preliminary finding suggested a nontoxic concentration of 55-60 µM for BAECs and BSMCs that could inhibit cell proliferation. This result may serve as the first step in assessing the use of honokiol in drug-eluting stents. MitoTracker Red was also applied to monitor the real-time effects of thrombectomy devices in live-cell artificial vessels. Four clot retrieval devices for stroke, Separator 3D, Trevo Pro Retrival System, Solitaire and Merci were separately tested in the artificial vessels lined with live endothelial cells that were developed for this study. The degrees of damages induced by the devices were revealed by MitoTracker Red staining. The results validated the application of live-cell artificial vessels in the study of endovascular devices. This thesis work identified MitoTracker Red as the most suitable multi-day live-cell tracking agent among several. Its applications then produced meaningful results in assessing the effects of drugs and thrombectomy devices

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