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首页> 外文期刊>Stem Cell Research & Therapy >Exosomes as potential alternatives to stem cell therapy for intervertebral disc degeneration: in-vitro study on exosomes in interaction of nucleus pulposus cells and bone marrow mesenchymal stem cells
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Exosomes as potential alternatives to stem cell therapy for intervertebral disc degeneration: in-vitro study on exosomes in interaction of nucleus pulposus cells and bone marrow mesenchymal stem cells

机译：外泌体作为干细胞治疗椎间盘退变的潜在替代方法：体外研究髓核细胞与骨髓间充质干细胞相互作用的外泌体

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Background The stem cell-based therapies for intervertebral disc degeneration have been widely studied. However, the mechanisms of mesenchymal stem cells interacting with intervertebral disc cells, such as nucleus pulposus cells (NPCs), remain unknown. Exosomes as a vital paracrine mechanism in cell–cell communication have been highly focused on. The purpose of this study was to detect the role of exosomes derived from bone marrow mesenchymal stem cells (BM-MSCs) and NPCs in their interaction with corresponding cells. Methods The exosomes secreted by BM-MSCs and NPCs were purified by differential centrifugation and identified by transmission electron microscope and immunoblot analysis of exosomal marker proteins. Fluorescence confocal microscopy was used to examine the uptake of exosomes by recipient cells. The effects of NPC exosomes on the migration and differentiation of BM-MSCs were determined by transwell migration assays and quantitative RT-PCR analysis of NPC phenotypic genes. Western blot analysis was performed to examine proteins such as aggrecan, sox-9, collagen II and hif-1α in the induced BM-MSCs. Proliferation and the gene expression profile of NPCs induced by BM-MSC exosomes were measured by Cell Counting Kit-8 and qRT-PCR analysis, respectively. Results Both the NPCs and BM-MSCs secreted exosomes, and these exosomes underwent uptake by the corresponding cells. NPC-derived exosomes promoted BM-MSC migration and induced BM-MSC differentiation to a nucleus pulposus-like phenotype. BM-MSC-derived exosomes promoted NPC proliferation and healthier extracellular matrix production in the degenerate NPCs. Conclusion Our study indicates that the exosomes act as an important vehicle in information exchange between BM-MSCs and NPCs. Given a variety of functions and multiple advantages, exosomes alone or loaded with specific genes and drugs would be an appropriate option in a cell-free therapy strategy for intervertebral disc degeneration.
机译：背景技术已经广泛研究了基于干细胞的椎间盘退变疗法。但是，间充质干细胞与椎间盘细胞（如髓核细胞（NPC））相互作用的机制仍然未知。外来体是细胞间通讯中重要的旁分泌机制。这项研究的目的是检测来自骨髓间充质干细胞（BM-MSC）和NPC的外泌体在与相应细胞相互作用中的作用。方法采用差速离心法纯化BM-MSCs和NPCs分泌的外泌体，并通过透射电镜和外泌体标记蛋白的免疫印迹分析进行鉴定。使用荧光共聚焦显微镜检查受体细胞对外泌体的摄取。 NPC外泌体对BM-MSCs迁移和分化的影响通过Transwell迁移测定和NPC表型基因的定量RT-PCR分析来确定。进行蛋白质印迹分析以检查诱导的BM-MSC中的蛋白，例如聚集蛋白聚糖，sox-9，胶原蛋白II和hif-1α。分别通过Cell Counting Kit-8和qRT-PCR分析测量了BM-MSC外泌体诱导的NPCs的增殖和基因表达谱。结果NPCs和BM-MSCs都分泌外泌体，这些外泌体被相应的细胞摄取。 NPC衍生的外泌体促进BM-MSC迁移并诱导BM-MSC分化为髓核样表型。 BM-MSC衍生的外来体在退化的NPC中促进NPC增殖和更健康的细胞外基质产生。结论我们的研究表明，外来体在BM-MSC和NPC之间的信息交换中起着重要的作用。考虑到多种功能和多种优势，在无细胞治疗椎间盘退变的策略中，单独或装载特定基因和药物的外泌体将是适当的选择。

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