Isolation and culture of rat embryonic neural cells: a quick protocol.

Isolation and culture of rat embryonic neural cells: a quick protocol.
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DOI:
10.3791/3965
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发表时间:
2012-05-24
期刊:
Journal of visualized experiments : JoVE
影响因子:
--
通讯作者:
Peruzzi F
Peruzzi F
中科院分区:
其他
文献类型:
--
作者:
Pacifici M;Peruzzi F

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我们描述了一种快速分离和培养E15-17大鼠胚胎海马或皮层神经元的方法。该方法可成功地应用于小鼠和人的原代神经元和神经祖细胞的分离。分离的神经元在无血清培养基中维持长达数周。这些培养物可用于核转染、免疫细胞化学、核酸制备以及电生理学。较老的神经元培养物也可以通过慢病毒转导以良好的效率转染,并且效率较低,用磷酸钙或基于脂质的方法如lipofectamine。
We are describing a quick method to dissociate and culture hippocampal or cortical neurons from E15-17 rat embryos. The procedure can be applied successfully to the isolation of mouse and human primary neurons and neural progenitors. Dissociated neurons are maintained in serum-free medium up to several weeks. These cultures can be used for nucleofection, immunocytochemistry, nucleic acids preparation, as well as electrophysiology. Older neuronal cultures can also be transfected with a good efficiency rate by lentiviral transduction and, less efficiently, with calcium phosphate or lipid-based methods such as lipofectamine.
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