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(all sterile filtered through 0.22 micron
filter)
- MEF Medium : 90% DMEM
(Dulbecco’s Modified Eagle Medium), 10% FBS (Fetal Bovine Serum
– Heat Inactivated), 1% glutamine, 1% NEAAs (non-essential amino
acids) (store at 4 oC)
- hESC Medium : 90% DMEM/F12
(Dulbecco’s Modified Eagle Medium/Ham’s F12 Mixture 1:1),
20% KnockOut Serum Replacement, 1% glutamine, 1% NEAAs (non-essential
amino acids) *(“complete medium” contains 10 ng/ml of
recombinant human basic fibroblast growth factor) (store at 4 oC)
- Mitomycin C Solution : 2 mg
Mitomycin C in 200 milliliters DMEM (Dulbecco’s Modified Eagle
Medium) (store in frozen aliquots)
- Collagenase IV Solution : 1.5 mg/ml
dissolved in DMEM/F12 (Dulbecco’s Modified Eagle
Medium/Ham’s F12 Mixture 1:1) (made fresh each time)
- hESC Freezing Medium (2x) : 60%
DMEM/F12 (Dulbecco’s Modified Eagle Medium/Ham’s F12
Mixture 1:1), 20% FBS (Fetal Bovine Serum – Heat Inactivated),
20% DMSO (Dimethyl Sulfoxide) (store at 4 oC)
- MEF Freezing Medium (2x) : 60% DMEM
(Dulbecco’s Modified Eagle Medium), 20% FBS (Fetal Bovine Serum
– Heat Inactivated), 20% DMSO (Dimethyl Sulfoxide) (store at 4 oC)
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