Purification of rat and mouse astrocytes by immunopanning.
Purification of rat and mouse astrocytes by immunopanning.
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DOI：
10.1101/pdb.prot074211
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发表时间：
2013-05-01
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通讯作者：
Foo, Lynette C
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作者：
Foo, Lynette C
我们描述了免疫泛素在纯化啮齿动物星形胶质细胞中的应用。星形胶质细胞的免疫泛化允许将星形胶质细胞从啮齿类动物的脑中分离出来。前瞻性分离指的是直接选择细胞，不需要多个步骤，持续几天，从而允许从皮质中选择具有代表性的比例的星形胶质细胞。由于免疫泛化是一个非常温和的过程，细胞在准备结束时是活的，可以在含有肝素结合的EGF样生长因子(HBEGF)的无血清介质中培养至少2周。HBEGF是星形胶质细胞在体外的关键生存因子。这个方案最初是在大鼠身上建立和验证的，但这里也描述了对小鼠细胞纯化的修改。
We describe the use of immunopanning to purify rodent astrocytes. Immunopanning of astrocytes permits the prospective isolation of astrocytes from the rodent brain. Prospective isolation refers to the direct selection of cells without multiple steps that extend over a few days, thereby permitting the selection of a representative proportion of the astrocytes from the cortex. Because immunopanning is a very gentle process, the cells are viable at the end of the preparation and can be cultured in a serum-free medium containing heparin-binding EGF-like growth factor (HBEGF), the critical survival factor of astrocytes in vitro, for at least 2 wk. This protocol was initially established and verified with rats, but modifications for the purification of mouse cells are also described here.