Grieco Giulio, Piccolo Simona, Ragni Enrico, de Girolamo Laura
Authors contributed equally.
Laboratorio di Biotecnologie Applicate all'Ortopedia, IRCCS Ospedale Galeazzi - Sant'Ambrogio, Milano I-20157, Italy.
Extracell Vesicles Circ Nucl Acids. 2025 Apr 29;6(2):195-215. doi: 10.20517/evcna.2024.99. eCollection 2025.
Mesenchymal stem cells (MSCs) are a promising therapeutic strategy for osteoarthritis (OA), largely due to their regenerative potential, which is attributed in part to their secretome. The secretome includes soluble factors and extracellular vesicles (EVs). Given that MSCs are sensitive to various culture conditions, this study aims to investigate the effects of different media supplemented with either fetal bovine serum (FBS) (F), platelet lysate (P), or serum/xeno-free (S/X) on the composition and therapeutic potential of the secretome from bone marrow-derived MSCs (BMSCs). BMSCs were cultured in F, P, or S/X media, with secretomes collected after starvation. The secretomes were analyzed for soluble factors, EVs, and miRNAs. Inflammatory responses were assessed in an OA model using inflamed chondrocytes and gene expression was evaluated by qRT-PCR. The secretomes from all conditions exhibited a similar molecular fingerprint. Proteomic analysis identified 98 common proteins encompassing growth factors and inflammatory mediators. EVs showed similar size and phenotype, with a slight difference in CD44 expression in EVs derived from P-expanded MSCs. Despite the high overall similarity, miRNA profiling identified 13 key players, with subtle differences in the miRNA composition of EVs from FBS-expanded BMSCs. All secretomes exhibited anti-inflammatory effects, with the FBS-expanded secretome showing the most pronounced therapeutic potential. The secretomes derived from different culture conditions share key molecular components. EVs may contribute to variations in therapeutic outcomes through their cargo. Optimizing MSC expansion conditions is crucial for enhancing the therapeutic potential of MSC-derived secretomes in OA treatment. Further research is needed to clarify the specific role of factors, miRNAs, and EVs in modulating OA pathology.
间充质干细胞（MSCs）是骨关节炎（OA）一种很有前景的治疗策略，这很大程度上归功于它们的再生潜力，而这种潜力部分归因于它们的分泌组。分泌组包括可溶性因子和细胞外囊泡（EVs）。鉴于MSCs对各种培养条件敏感，本研究旨在探讨添加胎牛血清（FBS）（F）、血小板裂解液（P）或无血清/无动物源成分（S/X）的不同培养基对骨髓来源的间充质干细胞（BMSCs）分泌组的组成和治疗潜力的影响。BMSCs在F、P或S/X培养基中培养，饥饿后收集分泌组。对分泌组的可溶性因子、EVs和微小RNA（miRNAs）进行分析。在OA模型中使用炎症软骨细胞评估炎症反应，并通过定量逆转录聚合酶链反应（qRT-PCR）评估基因表达。所有条件下的分泌组都表现出相似的分子指纹。蛋白质组学分析鉴定出98种常见蛋白质，包括生长因子和炎症介质。EVs显示出相似的大小和表型，来自P扩增的MSCs的EVs中CD44表达略有差异。尽管总体相似度很高，但miRNA谱分析鉴定出13个关键分子，来自FBS扩增的BMSCs的EVs的miRNA组成存在细微差异。所有分泌组均表现出抗炎作用，FBS扩增的分泌组显示出最显著的治疗潜力。不同培养条件下产生的分泌组共享关键分子成分。EVs可能通过其携带的物质导致治疗结果的差异。优化MSCs扩增条件对于提高MSCs来源的分泌组在OA治疗中的治疗潜力至关重要。需要进一步研究以阐明因子、miRNAs和EVs在调节OA病理中的具体作用。
