Soltero-Rivera Maria, Arzi Boaz, Bourebaba Lynda, Marycz Krzysztof
Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California-Davis, Davis, CA, United States.
Veterinary Institute for Regenerative Cures, School of Veterinary Medicine, University of California-Davis, Davis, CA, United States.
Stem Cells. 2025 Jun 24;43(7). doi: 10.1093/stmcls/sxaf014.
Extracellular vesicles (EVs) derived from mesenchymal stem cells have shown promise in treating inflammation. This study investigates whether preconditioning feline adipose-derived stem cells (FeASCs) with inflammatory cytokines, specifically IFN-γ and TNF-α, enhances the anti-inflammatory efficacy of MSC-derived EVs.
We hypothesize that cytokine-primed FeASCs will produce EVs with improved anti-inflammatory properties and that this preconditioning will affect mitochondrial dynamics to enhance EV therapy effectiveness.
FeASCs were exposed to a TNF-α/IFN-γ combination to mimic a pro-inflammatory milieu favoring ASCs' immunosuppressive phenotype. We analyzed morphological, metabolic, and immunomodulatory characteristics of native and cytokine-primed FeASCs. EVs were assessed for anti-inflammatory and mitochondrial-related markers. We also evaluated mitochondrial function and apoptosis markers in cytokine-primed cells.
Cytokine priming led to significant morphological changes in FeASCs, including enhanced cell projections and increased apoptosis. EVs from cytokine-primed FeASCs exhibited a heightened immunomodulatory profile, with increased expression of both pro-inflammatory and anti-inflammatory mediators. Transcriptomic analysis of these EVs revealed the upregulation of genes associated with cell proliferation, survival, and apoptosis. Mitochondrial function was impaired in cytokine-primed cells, but mitochondrial morphology remained unchanged. EVs from these cells contained higher levels of mitochondrial-related transcripts, indicating a compensatory response.
Cytokine-primed FeASCs generate EVs with enhanced immunomodulatory potential, highlighting their therapeutic promise. However, further research is needed to validate their efficacy and safety and refine preconditioning strategies to optimize EV-based therapies for inflammatory conditions. These advancements could pave the way for broader applications in regenerative medicine.
间充质干细胞衍生的细胞外囊泡（EVs）在治疗炎症方面已显示出前景。本研究调查用炎性细胞因子，特别是干扰素-γ（IFN-γ）和肿瘤坏死因子-α（TNF-α）预处理猫脂肪来源干细胞（FeASCs）是否会增强间充质干细胞衍生的EVs的抗炎功效。
我们假设细胞因子预处理的FeASCs将产生具有改善的抗炎特性的EVs，并且这种预处理将影响线粒体动力学以增强EV治疗效果。
将FeASCs暴露于TNF-α/IFN-γ组合以模拟有利于ASC免疫抑制表型的促炎环境。我们分析了天然和细胞因子预处理的FeASCs的形态、代谢和免疫调节特征。评估了EVs的抗炎和线粒体相关标志物。我们还评估了细胞因子预处理细胞中的线粒体功能和凋亡标志物。
细胞因子预处理导致FeASCs发生显著的形态变化，包括增强的细胞突起和凋亡增加。细胞因子预处理的FeASCs产生的EVs表现出更高的免疫调节特征，促炎和抗炎介质的表达均增加。对这些EVs的转录组分析揭示了与细胞增殖、存活和凋亡相关基因的上调。细胞因子预处理的细胞中线粒体功能受损，但线粒体形态保持不变。这些细胞产生的EVs含有更高水平的线粒体相关转录本，表明存在补偿反应。
细胞因子预处理的FeASCs产生具有增强免疫调节潜力的EVs，突出了它们的治疗前景。然而，需要进一步研究来验证它们的疗效和安全性，并完善预处理策略以优化基于EVs的炎症性疾病治疗。这些进展可为再生医学的更广泛应用铺平道路。
