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• W2014593046 abstract "Background: Measurement of vascular cell proliferation in animal models of hypertension is currently accomplished by demonstrating [3H]-thymidine ([3H]-dT) incorporation into DNA using autoradiography. This method, however, is labor intensive, requires radioactivity, and is limited by the inherent difficulty in discriminating labeled and unlabeled cells. To address these limitations, a flow cytometric-based method is described utilizing incorporation of 5-bromo-2′-deoxyuridine (BrdU) into DNA of nuclei isolated from blood vessels. Methods: Pulmonary hypertension was induced in rats by exposure to 10% O2 (hypoxia) for varying periods of time. Pulmonary arteries and aorta from rats injected with BrdU prior to sacrifice were isolated, fixed with 10% formalin, and digested with Protease XIV. The intact nuclei liberated by this treatment were successively treated with HCl/Triton X-100 and sodium borate. Processed nuclei were probed with a BrdU-specific fluorescein-conjugated antibody, and the percentage of BrdU staining cells was determined using flow cytometry. Results: An ≈20-fold increase in BrdU-positive cells at 3 days of hypoxia in pulmonary arteries (relative to control) with no change in aorta was observed. These results were similar to previous studies using [3H]-dT labeling. Conclusions: Flow cytometric determination of cell proliferation in blood vessels is a simple, objective technique that may facilitate measurement of cell proliferation in animal models of vascular disease. Cytometry 37:81–84, 1999. © 1999 Wiley-Liss, Inc." @default.
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• W2014593046 date "1999-09-01" @default.
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• W2014593046 title "Flow cytometric determination of cell proliferation in hypertensive blood vessels" @default.
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• W2014593046 doi "https://doi.org/10.1002/(sici)1097-0320(19990901)37:1<81::aid-cyto10>3.0.co;2-j" @default.
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