http://crutcherlab.med.uc.edu/DCN.htm

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