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细胞技术

Human Embryonic Stem (ES) Cell Protocols――Media and Reagents

2025-03-03 细胞技术 加入收藏
Serum Free Media for human ES cells on MEFs: can last for 7-10 daysFinal Conce

Serum Free Media for human ES cells on MEFs:   can last for 7-10 days

Final ConcentrationAmount for 250ml Stock solution
80% DMEM-F12200ml
20% KO Serum Replacer50ml
1% Non-essential Amino Acids2.5ml
1mM L-glutamine (see recipe)2.5ml
0.1mM b-mercaptoethanolpart of L-glutamine
4ng/ml bFGF (see recipe)0.5ml

L-glutamine stock solution:  Prepare on a per-use basis

  • 0.146g L-glutamine
  • 10ml Ca/Mg free PBS
  • 7 ul b-mercaptoethanol

bFGF Stock:  Store aliquots at �20 or �70

  • 10ug vial of bFGF
  • 5ml 0.1% Fraction V BSA
  • Aliquot 0.5ml into sterile tubes

0.1% Fraction V BSA:

  • 0.1g Fraction V BSA
  • 100ml PBS with Ca and Mg
  • Filter Sterilize

Collagenase IV Split Media:  This media lasts 2-3 weeks

  • Final concentration of 1mg/ml in DMEM-F12 media  ie. 0.05g collagenase IV in 50ml DMEM-F12

Cell Freezing Media:

Cryopreservative Medium:

Final ConcentrationAmount for 10ml Stock Solution
60% DMEM-F126ml
20% defined FBS2ml
20% DMSO (do not filter sterilize)2ml

 

Resuspension Medium:

Final ConcentrationAmount for 10ml Stock Solution
80% DMEM-F128ml
20% defined FBS2ml

 


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