Isolation of intact astrocytes from the optic nerve head of adult mice
Isolation of intact astrocytes from the optic nerve head of adult mice
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DOI：
10.1016/j.exer.2015.06.014
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发表时间：
2015-08-01
影响因子：
3.4
通讯作者：
Jakobs, Tatjana C.
中科院分区：
文献类型：
--
作者：
Choi, Hee Joo;Sun, Daniel;Jakobs, Tatjana C.
视神经乳头的星形胶质细胞是形成无髓神经节细胞轴突的直接细胞环境的白色星形胶质细胞的特化亚型。由于它们可能参与青光眼，这些星形胶质细胞已成为研究的目标。由于视神经组织的异质性，其中还包含其他细胞类型，在某些情况下，可能需要对少量充分表征的星形胶质细胞或甚至单个细胞进行基因表达研究。在这里，我们描述了一个简单的方法来分离单个星形胶质细胞。该方法允许从成年小鼠视神经获得具有完整形态的星形胶质细胞，并减少了分离的星形胶质细胞被其他细胞类型污染。单个星形胶质细胞可以通过其形态识别并在显微镜控制下收集。整个过程可在2-3小时内完成。我们还讨论了下游应用，如多重单细胞PCR和定量PCR（qPCR）。(C)2015爱思唯尔有限公司版权所有。
The astrocytes of the optic nerve head are a specialized subtype of white matter astrocytes that form the direct cellular environment of the unmyelinated ganglion cell axons. Due to their potential involvement in glaucoma, these astrocytes have become a target of research. Due to the heterogeneity of the optic nerve tissue, which also contains other cell types, in some cases it may be desirable to conduct gene expression studies on small numbers of well-characterized astrocytes or even individual cells. Here, we describe a simple method to isolate individual astrocytes. This method permits obtaining astrocytes with intact morphology from the adult mouse optic nerve and reduces contamination of the isolated astrocytes by other cell types. Individual astrocytes can be recognized by their morphology and collected under microscopic control. The whole procedure can be completed in 2-3 h. We also discuss downstream applications like multiplex single-cell PCR and quantitative PCR (qPCR). (C) 2015 Elsevier Ltd. All rights reserved.