Structural and Functional Biology Department, State University of Campinas (UNICAMP), São Paulo, Brazil.
Pharmacology Department, State University of Campinas (UNICAMP), São Paulo, Brazil.
Cytotherapy. 2019 Apr;21(4):444-459. doi: 10.1016/j.jcyt.2019.01.005. Epub 2019 Mar 20.
Endothelial progenitor cells (EPCs) are circulating progenitor cells that can play an essential role in vascular remodelling. In this work, we compared the role of two EPCs cultivated with different mediums in the resolution of the arterial thrombus induced by FeCl lesion and in vessel re-endothelization in the mouse carotid artery.
Mice mononuclear cells were differentiated into EPCs using Dulbecco's Modified Eagle's Medium (DMEM) and vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF) and IGF (Insulin Growth Factor) called EPCs--M1) or with EGM2(endothelial growth medium) (media supplemented with growth factors from Lonza called (EPCs-M2) for 30days and characterized using flow cytometry. The animals received three EPC injections post-lesion, and we analyzed thrombosis time, vessel re-endothelization, metalloproteinases activities, eNOS (endothelial Nitric oxide synthase) presence and SDF-1(Stromal Derived Factor- 1) levels in circulation.
EPC-M1 presented a more immature progenitor profile than EPC-M2 cells. The injection of EPC-M1 prolonged the thrombosis time, and the treatment with the different EPCs increased eNOS expression and MMP2 (Metalloproteinase 2) activity and decreased SDF-1 in plasma. Only EPC-M1 treatment increased both MMP2 and MMP9 and reduced thrombus after 7days. Also, both EPCs decreased platelet aggregation in vitro.
EPCs-M1 were more efficient in all of the analyzed assays. EPCsM2 may be a more mature EPC, proliferating less and promoting a less significant matrix remodelling. EPCs can promote vascular remodelling by inhibiting thrombosis and stimulating vascular wall remodelling and the treatment with a more immature progenitor may be more efficient in this process.
内皮祖细胞（EPCs）是循环祖细胞，在血管重塑中起着至关重要的作用。在这项工作中，我们比较了使用两种不同培养基培养的两种 EPC 在 FeCl 损伤诱导的动脉血栓溶解和小鼠颈总动脉血管再内皮化中的作用。
使用 DMEM（Dulbecco 改良 Eagle 培养基）和血管内皮生长因子（VEGF）、成纤维细胞生长因子（FGF）和 IGF（胰岛素样生长因子）将小鼠单核细胞分化为 EPCs，称为 EPC-M1）或用 EGM2（内皮生长培养基）（Lonza 提供的生长因子补充的培养基）培养 30 天，并通过流式细胞术进行特征分析。动物在损伤后接受三次 EPC 注射，我们分析了血栓形成时间、血管再内皮化、金属蛋白酶活性、循环中 eNOS（内皮一氧化氮合酶）的存在和 SDF-1（基质衍生因子-1）水平。
EPC-M1 比 EPC-M2 细胞具有更不成熟的祖细胞特征。EPC-M1 的注射延长了血栓形成时间，并且用不同的 EPC 处理增加了 eNOS 表达和 MMP2（基质金属蛋白酶 2）活性，降低了血浆中的 SDF-1。只有 EPC-M1 治疗增加了 MMP2 和 MMP9 的活性，并在 7 天后减少了血栓。此外，两种 EPC 都减少了体外血小板聚集。
EPC-M1 在所有分析的测定中都更有效。EPC-M2 可能是一种更成熟的 EPC，增殖较少，促进基质重塑的作用不明显。EPC 可通过抑制血栓形成和刺激血管壁重塑来促进血管重塑，使用更不成熟的祖细胞可能更有效地促进这一过程。
