Background:

Extracellular vesicles (EVs) are derived from internal cellular compartments, and have potential as a diagnostic and therapeutic tool in degenerative disease associated with aging. Mesenchymal stem cells (MSCs) have become a promising tool for functional EVs production. This study investigated the efficacy of EVs and its effect on differentiation capacity.

Methods:

The characteristics of MSCs were evaluated by flow cytometry and stem cell differentiation analysis, and a production mode of functional EVs was scaled from MSCs. The concentration and size of EVs were quantitated by Nanoparticle Tracking Analysis (NTA). Western blot analysis was used to assess the protein expression of exosome-specific markers. The effects of MSC-derived EVs were assessed by chondrogenic and adipogenic differentiation analyses and histological observation.

Results:

The range of the particle size of adipose-derived stem cells (ADSCs)- and Wharton’s jelly -MSCs-derived EVs were from 130 to 150 nm as measured by NTA, which showed positive expression of exosomal markers. The chondrogenic induction ability was weakened in the absence of EVs in vitro. Interestingly, after EV administration, type II collagen, a major component in the cartilage extracellular matrix, was upregulated compared to the EV-free condition. Moreover, EVs decreased the lipid accumulation rate during adipogenic induction.

Conclusion:

The results indicated that the production model could facilitate production of effective EVs and further demonstrated the role of MSC-derived EVs in cell differentiation. MSC-derived EVs could be successfully used in cell-free therapy to guide chondrogenic differentiation of ADSC for future clinical applications in cartilage regeneration.

中文翻译：

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间充质干细胞分泌的细胞外囊泡参与软骨细胞的产生并减少干细胞分化过程中的脂肪生成

背景：

细胞外囊泡 (EV) 源自内部细胞区室，具有作为与衰老相关的退行性疾病的诊断和治疗工具的潜力。间充质干细胞 (MSC) 已成为功能性 EV 生产的有前途的工具。本研究调查了 EV 的功效及其对分化能力的影响。

方法：

通过流式细胞术和干细胞分化分析评估 MSC 的特性，并从 MSC 扩展功能性 EV 的生产模式。EV 的浓度和大小通过纳米粒子跟踪分析 (NTA) 进行定量。蛋白质印迹分析用于评估外泌体特异性标记物的蛋白质表达。通过软骨形成和脂肪形成分化分析和组织学观察评估 MSC 衍生的 EV 的作用。

结果：

通过 NTA 测量，脂肪来源的干细胞 (ADSC) 和沃顿氏胶质细胞来源的 EV 的粒径范围为 130 至 150 nm，这表明外泌体标记物呈阳性表达。在体外没有 EV 的情况下，软骨形成诱导能力减弱。有趣的是，在给予 EV 后，与无 EV 条件相比，II 型胶原蛋白（软骨细胞外基质中的主要成分）被上调。此外，EV 降低了脂肪形成诱导过程中的脂质积累率。

结论：

结果表明，该生产模型可以促进有效 EV 的生产，并进一步证明了 MSC 衍生的 EV 在细胞分化中的作用。MSC 衍生的 EV 可成功用于无细胞疗法，以指导 ADSC 的软骨分化，用于未来软骨再生的临床应用。