Extracellular vesicles derived from macrophage promote angiogenesis In vitro and accelerate new vasculature formation In vivo.,Experimental Cell Research

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Extracellular vesicles derived from macrophage promote angiogenesis In vitro and accelerate new vasculature formation In vivo.
Experimental Cell Research ( IF 3.7 ) Pub Date : 2020-06-17 , DOI: 10.1016/j.yexcr.2020.112146
Prakash Gangadaran ₁ , Ramya Lakshmi Rajendran ₂ , Ji Min Oh ₂ , Chae Moon Hong ₃ , Shin Young Jeong ₄ , Sang-Woo Lee ₄ , Jaetae Lee ₄ , Byeong-Cheol Ahn ₅

Affiliation

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 pro-angiogenic 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.

中文翻译：

源自巨噬细胞的细胞外囊泡在体外促进血管生成并在体内加速新的脉管系统形成。

背景

局部缺血是组织血液供应的部分或完全阻塞。细胞外囊泡（EVs）新兴为缺血性疾病的治疗工具。大多数基于EV的缺血疗法均基于各种干细胞。在这里，我们提出了一种替代细胞源，用于分离促血管生成的电动汽车。

方法

从小鼠巨噬细胞系（Raw 264.7）分离出EV。巨噬细胞衍生的电动汽车（MAC-EVs）的特征使用透射电子显微镜，纳米粒子跟踪分析和蛋白质印迹（WB）分析进行了评估。进行WB和qRT-PCR以鉴定MAC-EV中促血管生成的VEGF和Wnt3a蛋白和microRNA（miR-210，miR-126和miR-130a）。进行了体外和体内Matrigel塞测定法，以研究MAC-EV用于管（类血管）形成和新血管形成的能力，并通过组织学评估。

结果

MAC-EV对ALIX呈阳性，而对钙结合蛋白呈阴性，呈圆形，平均大小为189±65.1 nm。WB和qRT-PCR结果表明，MAC-EVs中的VEGF，Wnt3a和miR-130a比细胞中的丰富。MAC-EVs治疗导致体外内皮细胞增殖，迁移和管形成增加。体内测定结果显示，与对照组相比，MAC-EVs增加了小鼠Matrigel栓中新的和较大的血管的形成。