Dissociated Cell Culture - Recipes

Embryo Extract:

1.    Chill 200, 3 d embryos after removing from chorions.
2.    Rinse in 0.5% chilled bleach for 2 minutes and then in zero calcium Ringer for 2 minutes.
3.    Transfer to a Dounce homogenizer with a minimum of liquid and homogenize well.
4.    Resuspend in 1 ml L-15 supplemented with 0.3 mg/ml glutamine, 50 U/ml penicillin, 0.05 mg/ml streptomycin and 0.8 mM CaCl₂.
5.    Store at -20°C.

Growth medium:

L-15 (Sigma)

0.3 mg/ml glutamine

50 U/ml penicillin

0.05 mg/ml streptomycin

0.8 mM CaCl₂

10% embryo extract

3% fetal calf serum.

Filter through a syringe filter.