Protocol for mixed cortical striatal cell culture 

Add 500µl of poly-D-lysine solution to each well that will be used for plating

Allow to sit inside the hood for at least 30mins (this plate will stay under the hood until dissection is finished)

Set BME + supplement and BME+AraC serum in water bath

Add 1-2ml of BMS alone into 2 wells of a 24-well plate (not coated)

Add 4-5ml of BMS alone into 16mm culture dish

Place dish on ice and leave BME inside the hood

Dissect P0/P1 brains on ice in to dissection medium

Aspirate medium gently, add 1ml of enzyme and incubate for 20 minutes at 37 degrees

Note: It is better to cut tissue sample using scalpel to allow more area for optimal enzymatic digestion

Wash 3X with 1ml of room temp BME medium without supplement to wash out the enzyme

Add 2ml of medium to well

Gently pipette 15 times and transfer to 15ml tube

Wait ~1min for large chunks to settle

Carefully remove supernatant and transfer to fresh 15ml tube

Centrifuge: 300g, 4°C, 4min

Remove media and resuspend in 1ml of media + serum

Remove poly-d-lysine from 24-well dishes

Add 500µl of medium + serum to each well

Count cells: 5x10^5 cells/well

Plate 1:2 striatum:cortex

Incubate for 1hr then change media

Add GDNF to BME-(without serum; 1µl of GDNF in 2.5ml)

Incubate cells in a CO2 incubator