In vitro model of glial scarring around neuroelectrodes chronically implanted in the CNS
In vitro model of glial scarring around neuroelectrodes chronically implanted in the CNS
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DOI：
10.1016/j.biomaterials.2006.06.018
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发表时间：
2006-11-01
期刊：
影响因子：
14
通讯作者：
Reichert, W. Monty
中科院分区：
文献类型：
--
作者：
Polikov, Vadim S.;Block, Michelle L.;Reichert, W. Monty
一种新的体外神经胶质瘢痕模型是通过采用一种基于原代细胞的系统开发的，该系统以前用于研究神经退行性疾病的神经炎症过程。将Fischer 344大鼠胚胎第14天的中脑机械分离，并在聚d -赖氨酸包被的24孔板上培养成神经元、星形胶质细胞和小胶质细胞的融合层。用机械刮伤或异物放置（直径50 μ m的不锈钢微丝段）对培养物进行损伤，在6小时至10天的时间点进行固定，并进行免疫细胞化学评估。小胶质细胞在早期时间点侵入擦伤创面，7天后增生活化的星形胶质细胞重新填充创面。微丝的长期存在导致10天胶质瘢痕形成，小胶质细胞形成覆盖微丝的内层细胞，而星形胶质细胞通过含有上调GFAP的细胞过程网络包围小胶质核心。在擦伤早期和形成胶质瘢痕的星形胶质细胞过程中存在表达Vimentin的细胞和过程。尽管通过膜片钳记录确定神经元具有电活性，但培养物中的神经元并没有重新填充擦伤伤口，也没有对微丝做出反应。胶质细胞对不同损伤模式的反应的时间过程和相对位置与体内典型的反应密切相关，这证明了在体外功能测试平台的进一步发展。(c) 2006 Elsevier Ltd.版权所有。
A novel in vitro model of glial scarring was developed by adapting a primary cell-based system previously used for studying neuroinflammatory processes in neurodegenerative disease. Midbrains from embryonic day 14 Fischer 344 rats were mechanically dissociated and grown on poly-D-lysine coated 24 well plates to a confluent layer of neurons, astrocytes, and microglia. The culture was injured with either a mechanical scrape or foreign-body placement (segments of 50 mu m diameter stainless steel microwire), fixed at time points from 6 h to 10 days, and assessed by immunocytochemistry. Microglia invaded the scraped wound area at early time points and hypertrophied activated astrocytes repopulated the wound after 7 days. The chronic presence of microwire resulted in a glial scar forming at 10 days, with microglia forming an inner layer of cells coating the microwire, while astrocytes surrounded the microglial core with a network of cellular processes containing upregulated GFAP. Vimentin expressing cells and processes were present in the scrape at early times and within the astrocyte processes forming the glial scar. Neurons within the culture did not repopulate the scrape wound and did not respond to the microwire, although they were determined to be electrically active through patch clamp recording. The time course and relative positions of the glia in response to the different injury paradigms correlated well with stereotypical in vivo responses and warrant further work in the development of a functional in vitro test bed. (c) 2006 Elsevier Ltd. All rights reserved.