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首页> 外文期刊>Experimental Eye Research >Microglia derived IL-6 suppresses neurosphere generation from adult human retinal cell suspensions.
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Microglia derived IL-6 suppresses neurosphere generation from adult human retinal cell suspensions.

机译：小胶质细胞来源的IL-6抑制了成人视网膜细胞悬浮液的神经球生成。

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摘要

Following retinal degeneration or inflammation that disrupts tissue architecture, there is limited evidence of tissue regeneration, despite evidence of cells with progenitor properties in the adult human retina at all ages. With the prospect of tissue/cell transplantation, redressing homeostasis whilst overcoming glial barrier or gliosis remains key to successful graft versus host integration and functional recovery. Activated human retinal microglia (MG) secrete cytokines, including IL-6, which may suppress neurogenesis or cellular (photoreceptor) replacement. To investigate this hypothesis, adult human retinal explants were cultured in cytokine-conditioned media (TNFalpha, TGFbeta, LPS/IFNgamma) to activate microglia in situ. Following culture of retinal explants for 4 days, supernatant conditioned by resulting migrated microglia was collected after a further 3 days and fed to retinal cell suspensions (RCS). Neurosphere (NS) generation and survival analysis was performed after 7 and 14 days in culture, with or without addition of conditioned media and with or without concomitant IL-6 neutralisation. Neurosphere phenotype was analysed by immunohistochemistry and cell morphology. Migratory MG from retinal explants were activated (iNOS-positive) and expressed CD45, CD11b, and CD11c. LPS/IFNgamma-activated MG conditioned media (MG-CM) contained significant levels of IL-6 (1265 +/- 143) pg/ml, which inhibited neurosphere generation within RCS in the presence of optimal neurosphere generating N2-FGF2 culture medium. Neutralising IL-6 activity reinstated NS generation and the differentiation capacity was maintained in the spheres that formed. Even in the presence of high levels of IL-6, those few NS that did form demonstrated a capacity to differentiate. The data supports activated MG-derived IL-6 influence retinal cell turnover.
机译：视网膜变性或炎症破坏了组织结构后，尽管有证据表明所有年龄段的成年人视网膜中都有具有祖细胞特性的细胞，但组织再生的证据有限。随着组织/细胞移植的前景，在克服神经胶质屏障或神经胶质增生的同时，平衡体内平衡仍然是成功进行移植物抗宿主整合和功能恢复的关键。活化的人类视网膜小胶质细胞（MG）分泌细胞因子，包括IL-6，可抑制神经发生或细胞（感光细胞）替代。为了研究该假设，将成年人类视网膜外植体在细胞因子条件培养基（TNFalpha，TGFbeta，LPS / IFNgamma）中培养以原位激活小胶质细胞。视网膜外植体培养4天后，再经过3天后收集通过迁移的小胶质细胞调节的上清液，然后喂入视网膜细胞悬液（RCS）。培养7天和14天后，在添加或不添加条件培养基以及有无IL-6中和的情况下，进行神经球（NS）的生成和生存分析。通过免疫组织化学和细胞形态学分析神经球表型。来自视网膜外植体的迁移性MG被激活（iNOS阳性）并表达CD45，CD11b和CD11c。 LPS / IFNgamma活化的MG条件培养基（MG-CM）包含显着水平的IL-6（1265 +/- 143）pg / ml，在存在产生最佳神经球的N2-FGF2培养基的情况下，其抑制了RCS内神经球的产生。中和的IL-6活性恢复了NS的生成，并且在形成的球体中维持了分化能力。即使存在高水平的IL-6，那些很少形成的NS仍具有分化能力。数据支持激活的MG衍生的IL-6影响视网膜细胞更新。

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