mES cell growth medium

Recipe

Reagent	Amount	Source
DMEM, 1X high glucose (+ glucose, + glutamine, without sodium pyruvate)	500 mL	GIBCO/Invitrogen
Fetal bovine serum (FBS), embryonic stem (ES) cell-tested, heat inactivated^a	50 mL	Atlanta Biologicals
mES supplement	27 mL

For mES cell growth medium, combine the reagents listed above and sterilize the solution with a 0.2-μM filter.
For mES cell growth medium with leukemia inhibitory factor (LIF; Millipore), combine 100 mL of mES cell growth medium with 50 μL of 10 μg/mL LIF (final concentration = 5 ng/mL) in a sterile bottle.
^aHeat-inactivate the ES-tested FBS by incubating it in a water bath for 30 min at 56ºC. Aliquot 50 mL into sterile conical tubes and store at −80ºC.