LiberaseTM cell dissociation for axolotl/frog limb cells

Prepare Reagents

0.7x PBS

• Dilute 1x PBS to 0.7x with dH2O

-> Chill on ice before use.

Liberase TM solution

• 40uL Stock Liberase TM (100x, 26WU/mL)
• 3960uL 0.7xPBS

-> Chill on ice before use.

AMEM(0) (serum-free medium)

• 114.3mL F12:DMEM+Glutmax
• 1.5mL Sodium pyruvate (100mM)
• 1.5mL B27 supplement
• 1.5mL Insulin (1 mg/mL)
• 1.5mL MEM Non-Essential Amino Acid (NEAA)
• 1.5mL Penicillin/Streptomycin (10000 U/mL)
• 28.2mL Sterile ddH2O

-> Total 150mL, filter.

-> Chill on ice before use.

High-Serum AMEM (HS-AMEM)

• 125mL Minimum essential medium (MEM)
• 20mL Heat-inactivated fetal bovine serum (56 degree, 30 min)
• 2mL Insulin (1 mg/mL)
• 2mL Glutamine (200mM)
• 2mL Penicillin/Streptomycin (10000 U/mL)
• 49mL Sterile ddH2O

-> Total 200mL, filter.

-> Chill on ice before use.