Department of Chemical and Biomedical Engineering, Florida State University, Tallahassee, Florida, USA.
Broad Stem Cell Research Center, David Geffen School of Medicine, University of California-Los Angeles (UCLA), Los Angeles, CA, USA.
J Extracell Vesicles. 2022 Jun;11(6):e12235. doi: 10.1002/jev2.12235.
Human mesenchymal stem cell (hMSC) derived extracellular vesicles (EVs) have shown therapeutic potential in recent studies. However, the corresponding therapeutic components are largely unknown, and scale-up production of hMSC EVs is a major challenge for translational applications. In the current study, hMSCs were grown as 3D aggregates under wave motion to promote EV secretion. Results demonstrate that 3D hMSC aggregates promote activation of the endosomal sorting complexes required for transport (ESCRT)-dependent and -independent pathways. mRNA sequencing revealed global transcriptome alterations for 3D hMSC aggregates. Compared to 2D-hMSC-EVs, the quantity of 3D-hMSC-EVs was enhanced significantly (by 2-fold), with smaller sizes, higher miR-21 and miR-22 expression, and an altered protein cargo (e.g., upregulation of cytokines and anti-inflammatory factors) uncovered by proteomics analysis, possibly due to altered EV biogenesis. Functionally, 3D-hMSC-EVs rejuvenated senescent stem cells and exhibited enhanced immunomodulatory potentials. In summary, this study provides a promising strategy for scalable production of high-quality EVs from hMSCs with enhanced therapeutic potential.
人源间充质干细胞（hMSC）衍生的细胞外囊泡（EVs）在最近的研究中显示出了治疗潜力。然而，相应的治疗成分在很大程度上是未知的，并且 hMSC EVs 的规模化生产是转化应用的主要挑战。在本研究中，hMSCs 在波运动下生长为 3D 聚集体以促进 EV 分泌。结果表明，3D hMSC 聚集体促进了内体分选复合物必需的运输（ESCRT）依赖和非依赖途径的激活。mRNA 测序揭示了 3D hMSC 聚集体的全基因组转录组改变。与 2D-hMSC-EVs 相比，3D-hMSC-EVs 的数量显著增加（增加了 2 倍），尺寸更小，miR-21 和 miR-22 的表达更高，通过蛋白质组学分析揭示了改变的蛋白质组（例如，细胞因子和抗炎因子的上调），这可能是由于 EV 发生了改变。在功能上，3D-hMSC-EVs 使衰老的干细胞恢复活力，并表现出增强的免疫调节潜力。总之，本研究提供了一种有前途的策略，可从 hMSCs 规模化生产具有增强治疗潜力的高质量 EVs。
