Microglia derived IL-6 suppresses neurosphere generation from adult human retinal cell suspensions
Microglia derived IL-6 suppresses neurosphere generation from adult human retinal cell suspensions
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DOI：
10.1016/j.exer.2009.06.019
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发表时间：
2009-11-01
影响因子：
3.4
通讯作者：
Dick, Andrew D.
中科院分区：
文献类型：
--
作者：
Balasubramaniam, Balini;Carter, Debra A.;Dick, Andrew D.
在视网膜变性或炎症破坏组织结构后，组织再生的证据有限，尽管在所有年龄的成年人视网膜中都有具有祖细胞特性的证据。随着组织/细胞移植的前景，在克服胶质屏障或胶质瘤的同时纠正体内平衡仍然是移植物与宿主成功整合和功能恢复的关键。激活的人视网膜小胶质细胞（MG）分泌细胞因子，包括IL-6，可能抑制神经发生或细胞（光感受器）替代。为了验证这一假设，我们在细胞因子条件培养基（TNF α、TGF β、LPS/IFN γ）中培养成人视网膜外植体，原位激活小胶质细胞。视网膜外植体培养4天后，3天后收集经迁移小胶质细胞调节的上清，并饲喂视网膜细胞悬液（RCS）。在培养7天和14天后进行神经球（NS）生成和生存分析，有或没有添加条件培养基，有或没有伴随IL-6中和。免疫组化和细胞形态学分析神经球表型。来自视网膜外植体的迁移MG被激活（inos阳性）并表达CD45， CD11b和CD11c。LPS/IFN激活MG条件培养基（MG- cm）含有显著水平的IL-6 (1265 +/- 143) pg/ml，在最佳神经球生成N2-FGF2培养基存在的情况下，抑制RCS内神经球的生成。中和的IL-6活性恢复了NS的生成，并维持了形成的球的分化能力。即使在高水平IL-6存在的情况下，那些少数形成的神经细胞也表现出分化的能力。数据支持活化的mg源性IL-6影响视网膜细胞更新。2009爱思唯尔有限公司版权所有。
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 (TNF alpha, TGF beta, LPS/IFN gamma) 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/IFN gamma-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. (C) 2009 Elsevier Ltd. All rights reserved.