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Differentiation of lymphatic endothelial cells from bone marrow mesenchymal stem cells with VEGFS

机译:VEGFS诱导骨髓间充质干细胞分化为淋巴管内皮细胞

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Although there have been many experimental studies demonstrating that bone marrow-derived mesenchymal stem cells (MSCs) have the potential to differentiate into mesenchymal tissues such as osteocytes, chondrocytes, and adipocytes in vivo and in vitro, little information is available regarding their potential to differentiate into lymphatic endothelial cells. Therefore, we chose to investigate differentiation of MSCs into lymphatic endothelial cells using stimulation with members of the vascular endothelial growth factor (VEGFs) family. Rat MSCs were isolated from bone marrow aspirate of Sprague-Dawley rats as previously described and characterized with flow cytometry for surface markers CD14, CD34, CD29, and CD90. Purified MSCs were plated and cultured in the presence of VEGF-A, VEGF-C, or the combination of both for 10 days. We examined the cells for Prox-1 and LYVE-1 by immunocytochemistry, RT-PCR, and Western blot analysis. Results demonstrated that compared to controls, cell differentiated with VEGF-A, VEGF-C and VEGF-A+VEGF-C expressed Prox-1 and LYVE-1. Our results indicate that MSCs induced by VEGFs are capable of differentiating into lymphatic endothelial-like cells in vitro, and this response has the potential to make them attractive candidates for the development of autologous tissue grafts for future therapy.
机译:尽管有许多实验研究表明,骨髓间充质干细胞(MSC)可能在体内和体外分化为间充质组织,例如骨细胞,软骨细胞和脂肪细胞,但关于它们分化的潜力的信息很少进入淋巴管内皮细胞因此,我们选择使用血管内皮生长因子(VEGF)家族的刺激物来研究MSCs向淋巴管内皮细胞的分化。如先前所述,从Sprague-Dawley大鼠的骨髓抽吸物中分离大鼠MSC,并用流式细胞仪对表面标记CD14,CD34,CD29和CD90进行表征。将纯化的MSC铺板并在VEGF-A,VEGF-C或两者的组合存在下培养10天。我们通过免疫细胞化学,RT-PCR和蛋白质印迹分析检查了细胞中的Prox-1和LYVE-1。结果表明,与对照相比,用VEGF-A,VEGF-C和VEGF-A + VEGF-C分化的细胞表达Prox-1和LYVE-1。我们的结果表明,由VEGF诱导的MSCs能够在体外分化为淋巴样内皮样细胞,这种反应具有使它们成为开发自体组织移植物以用于未来治疗的诱人候选物的潜力。

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