PC-12 growing culture

Male

Polygonal

PC-12 (CLS catalog number 500311)

Nerve growth factor (NGF)

Catecholamines; dopamine; norepinephrine

RPMI 1640

Suspension cells: Remove cells from substrate by pipetting with fresh medium. To obtain single cells, pass the suspension several times through a 22 gauge needle and dispense into new flasks. Growing on collagen: To remove adherent cells, use the following standard protocol. Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10ml for T75 cell culture flasks). Add TrypleExpress (1-2ml per T25, 2.5ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at 37 °C for 10 minutes. Carefully resuspend the cells, the addition of medium is optional but not necessary, and dispense into new flasks which contain fresh medium.

1 x 10^4 cells/cm^2

After thawing, plate the cells at 5 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 48 hours.

100

228

145

174

402

182