{"title":"连续营养供应培养策略控制细胞外囊泡的多泡内体途径和抗光老化miRNA货物装载。","authors":"Lihao Chen, Weihan Xie, Keke Wu, Yuan Meng, Yijun He, Jiawei Cai, Yuan Jiang, Qi Zhao, Yixi Yang, Minru Zhang, Manping Lu, Shaozhang Lin, Lin Liang, Zhiyong Zhang","doi":"10.1177/20417314231197604","DOIUrl":null,"url":null,"abstract":"<p><p>Extracellular vesicle (EV) therapy recently had shown significant efficacy in various diseases. Serum starvation culture (SC) is one of the most widely used methods for collecting EVs. However, SC may cause inadvertent effects and eventually dampen the therapeutic potential of EVs. Therefore, we developed a novel method for EV collection, continuous nutrient supply culture (CC), which can provide an optimal condition for mesenchymal stem cells (MSCs) by continuously supplying essential nutrients to MSCs. By comparing with SC strategy, we revealed that CC could maintain CC-MSCs in a normal autophagy and apoptotic state, which reduced the shunting of EV precursors in cells and useless information material carried by EVs. In CC-MSCs, the expression levels of endosomal sorting complexes required for transport (ESCRT) and targeting GTPase27 (Rab27) were upregulated compared to those in SC-MSCs. Besides, we analyzed the membrane transport efficiency of EV formation, which demonstrated the CC strategy could promote the formation of EV precursors and the release of EVs. 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引用次数: 0
摘要
细胞外囊泡(EV)治疗近年来在多种疾病中显示出显著的疗效。血清饥饿培养(SC)是目前应用最广泛的ev采集方法之一。然而,SC可能会引起无意的影响,并最终抑制ev的治疗潜力。因此,我们开发了一种新的EV收集方法——连续营养供应培养(CC),通过持续向间充质干细胞(MSCs)提供必需的营养物质,为其提供最佳的生长条件。通过与SC策略的比较,我们发现CC可以维持CC- mscs处于正常的自噬和凋亡状态,从而减少细胞内EV前体的分流和EV携带的无用信息物质。在CC-MSCs中,与SC-MSCs相比,运输所需的内体分选复合物(ESCRT)和靶向GTPase27 (Rab27)的表达水平上调。此外,我们还分析了EV形成的膜运输效率,表明CC策略可以促进EV前体的形成和EV的释放。此外,miRNA分析显示cc - ev富含抗慢性炎症因子,可抑制核因子κ b (NF-κB)通路,减轻慢性炎症,有效修复皮肤光老化损伤。
Continuous nutrient supply culture strategy controls multivesicular endosomes pathway and anti-photo-aging miRNA cargo loading of extracellular vesicles.
Extracellular vesicle (EV) therapy recently had shown significant efficacy in various diseases. Serum starvation culture (SC) is one of the most widely used methods for collecting EVs. However, SC may cause inadvertent effects and eventually dampen the therapeutic potential of EVs. Therefore, we developed a novel method for EV collection, continuous nutrient supply culture (CC), which can provide an optimal condition for mesenchymal stem cells (MSCs) by continuously supplying essential nutrients to MSCs. By comparing with SC strategy, we revealed that CC could maintain CC-MSCs in a normal autophagy and apoptotic state, which reduced the shunting of EV precursors in cells and useless information material carried by EVs. In CC-MSCs, the expression levels of endosomal sorting complexes required for transport (ESCRT) and targeting GTPase27 (Rab27) were upregulated compared to those in SC-MSCs. Besides, we analyzed the membrane transport efficiency of EV formation, which demonstrated the CC strategy could promote the formation of EV precursors and the release of EVs. In addition, miRNA analysis revealed that CC-EVs were enriched with anti-chronic inflammatory factors, which could inhibit the nuclear factor kappa-B (NF-κB) pathway, mitigate chronic inflammation, and effectively repair skin photo-aging damage.
期刊介绍:
The Journal of Tissue Engineering (JTE) is a peer-reviewed, open-access journal dedicated to scientific research in the field of tissue engineering and its clinical applications. Our journal encompasses a wide range of interests, from the fundamental aspects of stem cells and progenitor cells, including their expansion to viable numbers, to an in-depth understanding of their differentiation processes. Join us in exploring the latest advancements in tissue engineering and its clinical translation.