In vitro characterization and angiogenic potential of quality-and-quantity mononuclear cell-derived extracellular vesicles

IF 2.3 Q3 Medicine

JVS-vascular science Pub Date : 2026-01-01 Epub Date: 2026-02-12 DOI:10.1016/j.jvssci.2026.100410

Suttipong Tianwattanatada MD , Nuttapol Chruewkamlow PhD , Narinee Srimark MSc , Ladawan Khowawisetsut PhD , Kovit Pattanapanyasat PhD , Chutipon Sathan-ard MSc , Pisol Senawongse DDS, PhD , Nuttawut Sermsathanasawadi MD, PhD

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Abstract

Background

This study evaluated the presence, production rate, and in vitro angiogenic properties of quality-and-quantity culture mononuclear cells-derived extracellular vesicles (QQMNC-EVs).

Methods

Peripheral blood from three healthy volunteers was processed into five groups: QQMNC-EVs, QQMNCs, peripheral blood mononuclear cell-derived EVs (PBMNC-EVs), CD34⁺-EVs, and late endothelial progenitor cell-derived EVs (late EPC-EVs). EVs were identified and characterized using nanoparticle tracking analysis, flow cytometry, western blot analysis, and electron microscopy. EV production rates were quantified, and angiogenesis was evaluated via tube length formation assay.

Results

QQMNC-EVs were identified and met established EVs criteria. QQMNCs produced the highest EVs yield. This was significantly higher than CD34⁺ cells (P = .044), late EPCs (P = .004), and PBMNCs (P = .046). At low concentration, QQMNC-EVs showed angiogenic effects comparable with CD34⁺-EVs and late EPC-EVs, significantly greater than PBMNC-EVs (P = .018). At high concentration, QQMNC-EVs produced the highest tube length formation, demonstrating a dose-dependent effect. This was significantly higher than CD34⁺-EVs (P = .018), late EPC-EVs (P = .011), and PBMNC-EVs (P = .002).

Conclusions

QQMNC-EVs were successfully identified and showed in vitro angiogenic efficacy, highlighting their potential as a novel therapeutic strategy for patients with no-option chronic limb-threatening ischemia.

Clinical Relevance

Extracellular vesicles (EVs) were successfully identified and characterized from QQMNCs and had the highest production rate among the four cell groups: QQMNCs, standard peripheral blood mononuclear cells, CD34⁺ cells, and late endothelial progenitor cells. Moreover, QQMNC-EVs produced markedly higher tube length formation than standard mononuclear cell-derived EVs. These findings in angiogenic efficacy in vitro highlight their potential as a novel optional therapeutic strategy. Further investigation may advance treatment options for patients with no-option chronic limb-threatening ischemia.

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单个核细胞来源的细胞外囊泡的质量和数量的体外表征和血管生成潜力

本研究评估了单个核细胞来源的细胞外囊泡（QQMNC-EVs）的存在、产量和体外血管生成特性。方法将3名健康志愿者的外周血分为5组：qqmnc - ev、qqmnc - ev、外周血单个核细胞衍生ev、CD34+- ev和晚期内皮祖细胞衍生ev。利用纳米颗粒跟踪分析、流式细胞术、western blot分析和电子显微镜对ev进行鉴定和表征。量化EV产率，并通过管长形成试验评估血管生成。结果鉴定出qqmnc - ev，符合所建立的ev标准。QQMNCs的电动汽车产量最高。这明显高于CD34+细胞（P = 0.044）、晚期EPCs （P = 0.004）和pbmnc （P = 0.046）。在低浓度下，qqmnc - ev的血管生成作用与CD34+- ev和晚期epc - ev相当，显著高于pbmnc - ev （P = 0.018）。在高浓度下，qqmnc - ev形成的管长最高，且呈剂量依赖性。这明显高于CD34+- ev （P = 0.018）、晚期epc - ev （P = 0.011）和pbmnc - ev （P = 0.002）。结论sqqmnc - evs已被成功鉴定，并显示出体外血管生成的功效，这表明sqqmnc - evs有可能成为一种治疗慢性肢体缺血的新策略。细胞外囊泡（EVs）在QQMNCs、标准外周血单个核细胞、CD34+细胞和晚期内皮祖细胞四种细胞组中产量最高。此外，qqmnc - ev的管长形成明显高于标准的单核细胞衍生ev。这些体外血管生成功效的发现突出了它们作为一种新的可选治疗策略的潜力。进一步的研究可能会为无选择的慢性肢体威胁缺血患者提供治疗方案。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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