Serum Free Media for human ES cells on MEFs: can last for 7-10 days
Final Concentration | Amount for 250ml Stock solution |
---|---|
80% DMEM-F12 | 200ml |
20% KO Serum Replacer | 50ml |
1% Non-essential Amino Acids | 2.5ml |
1mM L-glutamine (see recipe) | 2.5ml |
0.1mM b-mercaptoethanol | part of L-glutamine |
4ng/ml bFGF (see recipe) | 0.5ml |
0.146g L-glutamine
10ml Ca/Mg free PBS
7 ul b-mercaptoethanol
10ug vial of bFGF
5ml 0.1% Fraction V BSA
Aliquot 0.5ml into sterile tubes
0.1g Fraction V BSA
100ml PBS with Ca and Mg
Filter Sterilize
Final concentration of 1mg/ml in DMEM-F12 media ie. 0.05g collagenase IV in 50ml DMEM-F12
Cell Freezing Media:
Final Concentration | Amount for 10ml Stock Solution |
---|---|
60% DMEM-F12 | 6ml |
20% defined FBS | 2ml |
20% DMSO (do not filter sterilize) | 2ml |
Final Concentration | Amount for 10ml Stock Solution |
---|---|
80% DMEM-F12 | 8ml |
20% defined FBS | 2ml |