脑皮质细胞分离培养 Culture of Dissociated Cortical Neurons
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脑皮质细胞分离培养 Culture of Dissociated Cortical Neurons
1. Use 7 to 8 day chicken embryos
2. Dissect forebrain in Ham's F12 media, peel off all meninges and then separate cortex from basal ganglia
3. Dissociate neurons by incubating with 0.5% trypsin for 20 minutes at 37°C
4. Wash tissue with fetal calf serum for 5 minutes to end trypsinization
5. Dissociate tissue by gentle trituration with a flamed pipet
6. Wash 2x in F12 medium, transfer to Neurobasal Medium (NBM, Gibco)
7. Plate onto pretreated (poly-dl -ornithine or poly-l -lysine) culture dishes containing NBM or F12+ medium.
8. Culture for 2 to 3 days
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