Crossland Rachel E, Sanjurjo-Rodríguez Clara, Reis Monica, Dickinson Anne M, Jones Elena, Wang Xiao-Nong
Translational and Clinical Research Institute, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne NE1 7RU, UK.
Institute of Biomedical Research of A Coruña (INIBIC), Centre of Advanced Scientific Researches (CICA), Universidade da Coruña, 15006 A Coruña, Spain.
Int J Mol Sci. 2025 Jul 21;26(14):7010. doi: 10.3390/ijms26147010.
Mesenchymal stromal cells (MSCs) have demonstrated therapeutic efficacy across numerous clinical applications, with evidence suggesting their paracrine effects, particularly through extracellular vesicles (EVs), possibly driving functional outcomes. In this study we perform the comprehensive characterization of microRNA expression profiles in human MSC-derived EVs (MSC-EV) compared to their parental cells, cultured under clinically relevant xeno-free conditions. MSCs were isolated from the bone marrows of healthy donors and characterised according to the International Society for Cellular Therapy criteria, while MSC-EVs were isolated using differential ultracentrifugation and validated according to the International Society for Extracellular Vesicle guidelines. NanoString profiling identified 590 mature microRNAs expressed across both populations, with 42 being significantly differentially expressed between MSC-EVs and parental MSCs. Five microRNAs were distinctly highly expressed in MSCs and five in MSC-EVs, while fifteen of the top twenty most abundant microRNAs showed high expression in both populations. MicroRNA expression patterns were validated in an independent cohort. Functional pathway analysis of differentially expressed microRNAs showed enrichment of key biological processes including cell proliferation, differentiation, and immune regulation. This standardised profiling approach develops our understanding of MSC/MSC-EV microRNA cargo, using a transparent methodological approach that allows for the improved comparability of datasets for the development and advancement of MSC-EV therapeutics.
间充质基质细胞（MSCs）已在众多临床应用中展现出治疗效果，有证据表明其旁分泌作用，尤其是通过细胞外囊泡（EVs），可能推动了功能结果。在本研究中，我们对在临床相关无血清条件下培养的人MSC来源的EVs（MSC-EV）与其亲代细胞相比的微小RNA表达谱进行了全面表征。从健康供体的骨髓中分离出MSCs，并根据国际细胞治疗协会标准进行表征，而MSC-EVs则通过差速超速离心法分离，并根据国际细胞外囊泡协会指南进行验证。NanoString分析鉴定出在两个群体中均表达的590种成熟微小RNA，其中42种在MSC-EVs和亲代MSCs之间存在显著差异表达。5种微小RNA在MSCs中明显高表达，5种在MSC-EVs中高表达，而在最丰富的前20种微小RNA中，有15种在两个群体中均高表达。在一个独立队列中验证了微小RNA的表达模式。对差异表达微小RNA的功能通路分析显示，关键生物学过程包括细胞增殖、分化和免疫调节均有富集。这种标准化的分析方法加深了我们对MSC/MSC-EV微小RNA负载的理解，采用了一种透明的方法，可提高数据集的可比性，以促进MSC-EV治疗学的发展和进步。
