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鹌鹑胚胎视网膜器官型培养中微胶质细胞的迁移和分支。

Migration and ramification of microglia in quail embryo retina organotypic cultures.

机构信息

Departamento de Biología Celular, Facultad de Ciencias, Universidad de Granada, E-18071 Granada, Spain.

出版信息

Dev Neurobiol. 2011 Apr;71(4):296-315. doi: 10.1002/dneu.20860.

Abstract

Organotypic cultures of retina explants preserve the complex cellular microenvironment of the retina and have been used as a tool to assess the biological functions of some cell types. However, studies to date have shown that microglial cells activate quickly in response to the retina explantation. In this study, microglial cells migrated and ramified in quail embryo retina organotypic cultures (QEROCs) according to chronological patterns bearing a resemblance to those in the retina in situ, despite some differences in cell density and ramification degree. Retinal explants from quail embryos at 9 days of incubation (E9) proved to be the best in vitro system for reproducing a physiological-like behavior of microglial cells when cultured in Eagle's basal medium supplemented with horse serum. During the first week in vitro, microglial cells migrated tangentially in the vitreal part of QEROCs, and some began to migrate radially from 3 days in vitro (div) onward, ramifying in the inner and outer plexiform layers, thus mimicking microglia development in the retina in situ, although reaching a lower degree of ramification after 7 div. From 8 div onward, microglial cells rounded throughout the explant thickness simultaneously with the nonphysiological appearance of dead photoreceptors and round microglia in the outernuclear layer. Therefore, E9 QEROCs can be used during the first week in vitro as a model system for experimental studies of molecules putatively involved in microglial migration and ramification.

摘要

器官型培养的视网膜外植体保留了视网膜复杂的细胞微环境,已被用作评估某些细胞类型生物学功能的工具。然而,迄今为止的研究表明,小胶质细胞在外植体分离后会迅速激活。在这项研究中,小胶质细胞根据时间顺序在鹌鹑胚胎视网膜器官型培养物(QEROC)中迁移和分支,其模式与原位视网膜相似,尽管细胞密度和分支程度存在一些差异。孵育 9 天(E9)的鹌鹑胚胎的视网膜外植体在补充有马血清的 Eagle 基础培养基中培养时,被证明是体外再现小胶质细胞生理样行为的最佳系统。在体外培养的第一周,小胶质细胞在 QEROC 的玻璃体部分呈切线状迁移,一些小胶质细胞从第 3 天开始呈放射状迁移,在内外丛状层分支,从而模拟了原位视网膜中小胶质细胞的发育,尽管在 7 天后分支程度较低。从第 8 天开始,小胶质细胞在整个外植体厚度上同时变圆,同时出现非生理状态的死亡光感受器和外核层中的圆形小胶质细胞。因此,E9 的 QEROC 可以在体外培养的第一周内用作实验研究分子的模型系统,这些分子可能参与小胶质细胞的迁移和分支。

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