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Comparison between Culture Conditions Improving Growth and Differentiation of Blood and Bone Marrow Cells Committed to the Endothelial Cell Lineage

机译:促进内皮细胞谱系的血液和骨髓细胞生长和分化的培养条件的比较

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The aim of this study was to compare different cell sources and culture conditions to obtain endothelial progenitor cells (EPCs) with predictable antigen pattern, proliferation potential and in vitro vasculogenesis. Pig mononuclear cells were isolated from blood (PBMCs) and bone marrow (BMMCs). Mesenchymal stem cells (MSCs) were also derived from pig bone marrow. Cells were cultured on fibronectin in the presence of a high concentration of VEGF and low IGF-1 and FGF-2 levels, or on gelatin with a lower amount of VEGF and higher IGF-1 and FGF-2 concentrations. Endothelial commitment was relieved in almost all PBMCs and BMMCs irrespective of the protocol used, whilst MSCs did not express a reliable pattern of EPC markers under these conditions. BMMCs were more prone to expand on gelatin and showed a better viability than PBMCs. Moreover, about 90% of the BMMCs pre-cultured on gelatin could adhere to a hyaluronan-based scaffold and proliferate on it up to 3 days. Pre-treatment of BMMCs on fibronectin generated well-shaped tubular structures on Matrigel, whilst BMMCs exposed to the gelatin culture condition were less prone to form vessel-like structures. MSCs formed rough tubule-like structures, irrespective of the differentiating condition used. In a relative short time, pig BMMCs could be expanded on gelatin better than PBMCs, in the presence of a low amount of VEGF. BMMCs could better specialize for capillary formation in the presence of fibronectin and an elevated concentration of VEGF, whilst pig MSCs anyway showed a limited capability to differentiate into the endothelial cell lineage.
机译:这项研究的目的是比较不同的细胞来源和培养条件,以获得具有可预测的抗原模式,增殖潜力和体外血管生成的内皮祖细胞(EPC)。从血液(PBMC)和骨髓(BMMC)中分离出猪单核细胞。间充质干细胞(MSCs)也来源于猪骨髓。在存在高浓度VEGF和低IGF-1和FGF-2水平的纤连蛋白上培养细胞,或在具有较低VEGF量和较高IGF-1和FGF-2浓度的明胶上培养细胞。无论使用哪种方案,几乎所有PBMC和BMMC的内皮细胞承诺都得到了缓解,而MSC在这些条件下并未表达可靠的EPC标记模式。 BMMC比PBMC更易于在明胶上扩展,并显示出更好的生存能力。此外,在明胶上预培养的BMMC约有90%可以粘附在透明质酸基支架上并在其上增殖长达3天。在纤连蛋白上预处理BMMC在Matrigel上产生形状良好的管状结构,而暴露于明胶培养条件下的BMMC不易形成血管样结构。 MSC形成粗糙的肾小管样结构,与使用的分化条件无关。在存在少量VEGF的情况下,在相对较短的时间内,猪BMMCs可以比PBMCs更好地在明胶上扩增。在存在纤连蛋白和高浓度VEGF的情况下,BMMC可以更好地专门用于毛细血管的形成,而无论如何,猪MSC都显示出分化为内皮细胞谱系的能力有限。

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