Tina:LabNotes/2012-6-4
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- Collect MEF conditioned medium (2*10-2+36=54 mL), store at -20. Add new mESC medium to MEF cells.
- Passage feeder free P9 InsACGGsAlov3 mESCs into 1:6 (1 well, feeder free condition, Add 2 mL MEF conditioned medium).
- Thaw one vial of MITC treated MEF CF-1 cells (GlobalStem, Cat# GSC-6001M, Lot# 94300663) in one T75 and two wells of a 6-well plate. (Originally seeded in four T75, quickly transferred to current seeding density within 30 mins, might be some cell lose..)
- Prepare 500 mL mouse ES cell culture medium.(403.5 mL Knockout DMEM + 75 mL ES-FBS + 6 mL homemade LIF(from Dr. Xu's Lab) + 5 mL NEAA + 5 mL Glutamax + 5 mL Pen/Strep (expired) + 500 uL BME soultion)
- Passage P9 InsACGGsAlov3 mESCs into 1:6 (1 well, with MEF).