Song:MEF Preparation
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- Grow passage-3(no more than) primary MEFs to confluency.
- Remove the medium, add 10 μg/ml mitomycin-c (diluted in MEF medium) and incubate at 37 °C for 3 h.
- Remove the mitomycin-c solution from the cells and wash them with DPBS for five times.
- Trypsinize the cells, collect in MEF medium, and then centrifuge them.
- aliquot into cryovials and freeze at –80 degree.