Mellows Ben, Mitchell Robert, Antonioli Manuela, Kretz Oliver, Chambers David, Zeuner Marie-Theres, Denecke Bernd, Musante Luca, Ramachandra Durrgah L, Debacq-Chainiaux Florence, Holthofer Harry, Joch Barbara, Ray Steve, Widera Darius, David Anna L, Huber Tobias B, Dengjel Joern, De Coppi Paolo, Patel Ketan
1 School of Biological Sciences, University of Reading , Reading, United Kingdom .
2 Department of Epidemiology and Preclinical Research, National Institute for Infectious Diseases IRCCS 'Lazzaro Spallanzani' , Rome, Italy .
Stem Cells Dev. 2017 Sep 15;26(18):1316-1333. doi: 10.1089/scd.2017.0089. Epub 2017 Aug 22.
The secretome of human amniotic fluid stem cells (AFSCs) has great potential as a therapeutic agent in regenerative medicine. However, it must be produced in a clinically compliant manner before it can be used in humans. In this study, we developed a means of producing a biologically active secretome from AFSCs that is free of all exogenous molecules. We demonstrate that the full secretome is capable of promoting stem cell proliferation, migration, and protection of cells against senescence. Furthermore, it has significant anti-inflammatory properties. Most importantly, we show that it promotes tissue regeneration in a model of muscle damage. We then demonstrate that the secretome contains extracellular vesicles (EVs) that harbor much, but not all, of the biological activity of the whole secretome. Proteomic characterization of the EV and free secretome fraction shows the presence of numerous molecules specific to each fraction that could be key regulators of tissue regeneration. Intriguingly, we show that the EVs only contain miRNA and not mRNA. This suggests that tissue regeneration in the host is mediated by the action of EVs modifying existing, rather than imposing new, signaling pathways. The EVs harbor significant anti-inflammatory activity as well as promote angiogenesis, the latter may be the mechanistic explanation for their ability to promote muscle regeneration after cardiotoxin injury.
人羊水干细胞（AFSCs）的分泌组作为再生医学中的一种治疗剂具有巨大潜力。然而，在用于人体之前，它必须以符合临床规范的方式生产。在本研究中，我们开发了一种从AFSCs生产不含所有外源性分子的生物活性分泌组的方法。我们证明完整的分泌组能够促进干细胞增殖、迁移并保护细胞免于衰老。此外，它具有显著的抗炎特性。最重要的是，我们表明它在肌肉损伤模型中促进组织再生。然后我们证明分泌组包含细胞外囊泡（EVs），其具有整个分泌组的许多但不是全部生物活性。EV和游离分泌组分的蛋白质组学表征显示每个组分存在大量特定分子，这些分子可能是组织再生的关键调节因子。有趣的是，我们表明EVs仅包含miRNA而不包含mRNA。这表明宿主中的组织再生是由EVs修饰现有信号通路而非施加新信号通路的作用介导的。EVs具有显著的抗炎活性并促进血管生成，后者可能是其在心脏毒素损伤后促进肌肉再生能力的机制解释。
