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KMID : 0370220180620050313
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Yakhak Hoeji
2018 Volume.62 No. 5 p.313 ~ p.318
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Two-cell tube formation assay system for angiogenesis research
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Kang Kyu-Tae
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Abstract
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Blood vessels are formed by two cell types: endothelial cells and vascular mural cells. However, most in vitro angiogenesis assays are performed by only endothelial cells. Here, we developed an in vitro angiogenesis assay system using two human vascular cell precursors: endothelial colony forming cells (ECFC) and mesenchymal stem cells (MSC). The progression of tube formation was monitored by a real-time cell recorder for 24 hours. We also utilized an image-stitching software to increase the analyzed image area up to 9-times larger than one image from 10X object lens. Tube number was higher in ECFC+MSC than ECFC at 6 hour time point, but became lower at 12 and 24 hours when compared with ECFC. The length per tube was higher in ECFC+MSC than ECFC at all-time points. VEGF treatment increased tube formation in both ECFC+MSC and ECFC. Vatalanib, VEGF inhibitor, inhibited tube formation of ECFC dose-dependently. Interestingly, ECFC+MSC-induced tube formation was not changed by vatalanib, suggesting that MSC may potentiate the stability and durability of tubular structures. These results indicate that two-cell tube formation assay system can mimic in vivo angiogenic process by two vascular cells, and will be an effective pre-clinical in vitro assay model to evaluate pro- or anti-angiogenic property.
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KEYWORD
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lial colony forming cells, mesenchymal stem cells, angiogenesis, tube formation assay
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