Extracellular vesicles derived from macrophage promote angiogenesis In vitro and accelerate new vasculature formation In vivo
Extracellular vesicles derived from macrophage promote angiogenesis In vitro and accelerate new vasculature formation In vivo
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DOI：
10.1016/j.yexcr.2020.112146
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发表时间：
2020-09-15
影响因子：
3.7
通讯作者：
Ahn, Byeong-Cheol
中科院分区：
文献类型：
--
作者：
Gangadaran, Prakash;Rajendran, Ramya Lakshmi;Ahn, Byeong-Cheol
背景：缺血是指部分或完全阻断组织的血液供应。细胞外小泡(EVS)作为一种治疗缺血性疾病的工具正在兴起。大多数基于EV的缺血疗法都是基于各种干细胞的。在这里，我们提出了一种分离促血管生成血管内皮细胞的替代细胞来源。方法：从小鼠巨噬细胞系(RAW 264.7)分离EV。用透射电子显微镜、纳米颗粒跟踪分析和Western blotting(WB)分析评价巨噬细胞来源的EVS的特征。Wb和qRT-PCR检测MAC-EVS中促血管生成的VEGF、Wnt3a蛋白和microRNAs(miR-210、miR-126和miR-130a)。结果：MACEVS呈圆形，平均大小为189+/-65.1 nm，Alix阳性，Calnexin阴性。Wb和qRT-PCR结果显示，血管内皮生长因子、Wnt3a和miR-130a在MAC-EVS中比在细胞中表达更丰富。在体外，Mac-EVS处理可增加内皮细胞的增殖、迁移和管状形成。体内实验结果显示，与对照组相比，MAC-EVS可促进小鼠Matrigel栓内新生血管的形成。结论：MAC-EVS在体外和体内均具有诱导血管生成的潜能，可作为治疗缺血性疾病的促血管生成的替代物。
Background: Ischemia is the partial or complete blockage of blood supply to tissues. Extracellular vesicles (EVs) are emerging as a therapeutic tool for ischemic diseases. Most EV-based ischemia therapies are based on various stem cells. Here, we propose an alternative cell source for the isolation of pro-angiogenic EVs.Methods: EVs were isolated from a mouse macrophage cell line (Raw 264.7). The characteristic features of the macrophage-derived EVs (MAC-EVs) were assessed using transmission electron microscopy, nanoparticle tracking analysis, and Western blotting (WB) analysis. WB and qRT-PCR were performed to identify the proangiogenic VEGF and Wnt3a proteins and microRNAs (miR-210, miR-126, and miR-130a) in the MAC-EVs. In vitro and in vivo Matrigel plug assays were performed to investigate the capacity of the MAC-EVs for tube (blood vessel-like) formation and new blood vessel formation and assessed by histology.Results: The MAC-EVs was positive for ALIX and negative for calnexin, with a round shape and an average size of 189 +/- 65.1 nm. WB and qRT-PCR results revealed that VEGF, Wnt3a and miR-130a were more abundant in the MAC-EVs than cells. MAC-EVs treatment resulted in increased endothelial cellular proliferation, migration, and tube formation in vitro. In vivo assay results revealed that MAC-EVs increased the formation of new and larger blood vessels in the Matrigel plug of mice compared to the formation in the control group.Conclusion: Our results suggest that MAC-EVs have the potential to induce angiogenesis in vitro and in vivo, could serve as a pro-angiogenic alternative for ischemic diseases.