Isolation and Culture of Mouse Cortical Astrocytes
Isolation and Culture of Mouse Cortical Astrocytes
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DOI：
10.3791/50079
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发表时间：
2013-01-01
影响因子：
1.2
通讯作者：
Schachtrup, Christian
中科院分区：
文献类型：
--
作者：
Schildge, Sebastian;Bohrer, Christian;Schachtrup, Christian
星形胶质细胞是哺乳动物大脑中数量丰富的细胞类型，但其分子和功能特征仍有待进一步研究。体外星形胶质细胞培养系统可用于详细研究这些胶质细胞的生物学功能。本视频展示了如何通过分离和培养小鼠幼崽的混合皮层细胞来获得纯星形胶质细胞。该方法是基于缺乏活的神经元和星形胶质细胞，少突胶质细胞和小胶质细胞，三个主要的神经胶质细胞群体的中枢神经系统，在文化的分离。在培养的第一天期间的代表性图像证明了混合细胞群体的存在，并且指示了星形胶质细胞变得融合并且应当与小胶质细胞和少突胶质细胞分离的时间点。此外，我们证明纯度和星形胶质细胞形态培养星形胶质细胞使用免疫细胞化学染色以及建立和新描述的星形胶质细胞标志物。该培养体系可方便地获得纯化的小鼠星形胶质细胞和星形胶质细胞条件培养液，用于星形胶质细胞生物学的研究。
Astrocytes are an abundant cell type in the mammalian brain, yet much remains to be learned about their molecular and functional characteristics. In vitro astrocyte cell culture systems can be used to study the biological functions of these glial cells in detail. This video protocol shows how to obtain pure astrocytes by isolation and culture of mixed cortical cells of mouse pups. The method is based on the absence of viable neurons and the separation of astrocytes, oligodendrocytes and microglia, the three main glial cell populations of the central nervous system, in culture. Representative images during the first days of culture demonstrate the presence of a mixed cell population and indicate the timepoint, when astrocytes become confluent and should be separated from microglia and oligodendrocytes. Moreover, we demonstrate purity and astrocytic morphology of cultured astrocytes using immunocytochemical stainings for well established and newly described astrocyte markers. This culture system can be easily used to obtain pure mouse astrocytes and astrocyte-conditioned medium for studying various aspects of astrocyte biology.