Culture of Cortical Neurons from E18 Prenatal Rats
WANG Jinpeng, ZHANG Yong*
Primary neuronal culture from prenatal rats is widely used to uncover cellular mechanisms underlying various processes in neurons such as cellular trafficking, cellular structure and protein localization. However, unambiguous results from neuronal culture depend on the health, purity and complexity of the neurons. Here we provide a protocol for isolating and culturing cortical neurons from E18 embryonic rats. We discuss detailed culture techniques including glass coverslips treatment, cortex dissection, digestion, plating and medium replacement. Under our protocol, the cultured cortical neurons could be maintained in healthy condition (neuronal morphology, extensive axonal and dendritic arbors development) for at least 20 days. The cultured neurons were tested for immunostaining and confocal imaging, we can also achieve a 10% transfection efficiency with the neuronal culture. Overall, this optimized cell culture technique can be used to culture and maintain the normal development of neurons for applications such as immunohistochemistry, gene editing as well as live imaging studies.
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