Chen Yin-Hsueh, Duan Xu, Nie Qing, Li Li-Jun, Chen Gang, Li Ye, Yang Hong-Yu
Department of Oral and Maxillofacial Surgery, Stomatological Center, Peking University Shenzhen Hospital, Shenzhen 518000, China.
The State Key Laboratory of Oral & Maxillofacial Reconstruction and Regeneration, Key Laboratory of Oral Biomedicine Ministry of Education, Hubei Key Laboratory of Stomatology, School & Hospital of Stomatology, Wuhan University, Wuhan 430079, China.
ACS Biomater Sci Eng. 2025 Sep 8;11(9):5572-5585. doi: 10.1021/acsbiomaterials.5c00898. Epub 2025 Aug 27.
Extracellular vesicles (EVs) derived from mesenchymal stem cells (MSC-EVs) are promising therapeutic agents for various diseases. However, current methods to improve MSC-EV production are insufficient to meet the clinical demands. Although various strategies have been investigated to enhance MSC-EV production, they are often hampered by limited scalability, loss of stemness, or suboptimal therapeutic outcomes. Our study identified three key stimulators that significantly boosted MSC-EV production: epidermal growth factor (EGF), tumor necrosis factor-α (TNF-α), and hypoxia. Employing an orthogonal design, we developed an optimized cell culture condition, subsequently referred to as ETH (EGF 10 ng/mL, TNF-α 50 ng/mL, and a hypoxic environment of 1% O) preconditioning. This approach led to a remarkable 4- to 5-fold increase in MSC-EV yield while preserving the stemness of MSCs. Through proteomic analysis, we elucidated the underlying mechanisms of ETH preconditioning, providing insight into the complex processes driving enhanced MSC-EV production. Notably, MSC-EVs generated through ETH preconditioning demonstrated enhanced therapeutic potential including superior angiogenesis, collagen deposition, and regulation of inflammation. These findings present a scalable and effective strategy for elevating MSC-EV production, paving the way for its broader clinical application in regenerative medicine.
间充质干细胞衍生的细胞外囊泡（MSC-EV）是治疗多种疾病的有前景的治疗剂。然而，目前改善MSC-EV产量的方法不足以满足临床需求。尽管已经研究了各种提高MSC-EV产量的策略，但它们往往受到可扩展性有限、干性丧失或治疗效果欠佳的阻碍。我们的研究确定了三种显著提高MSC-EV产量的关键刺激物：表皮生长因子（EGF）、肿瘤坏死因子-α（TNF-α）和缺氧。采用正交设计，我们开发了一种优化的细胞培养条件，随后称为ETH（EGF 10 ng/mL、TNF-α 50 ng/mL和1% O的低氧环境）预处理。这种方法使MSC-EV产量显著提高了4至5倍，同时保留了MSC的干性。通过蛋白质组学分析，我们阐明了ETH预处理的潜在机制，深入了解了驱动MSC-EV产量增加的复杂过程。值得注意的是，通过ETH预处理产生的MSC-EV显示出增强的治疗潜力，包括卓越的血管生成、胶原蛋白沉积和炎症调节。这些发现提出了一种提高MSC-EV产量的可扩展且有效的策略，为其在再生医学中更广泛的临床应用铺平了道路。
