Song:MEF Preparation: Difference between revisions
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#Trypsinize the cells, collect in MEF medium, and then centrifuge. | #Trypsinize the cells, collect in MEF medium, and then centrifuge. | ||
#Aliquot into cryovials and freeze at –80 °C. | #Aliquot into cryovials and freeze at –80 °C. | ||
#Refs:[http://www.lifetechnologies.com/us/en/home/references/protocols/cell-culture/stem-cell-protocols/ipsc-protocols/feeder-dependent-culture-hescs.html Cell number] | #Refs:[http://www.lifetechnologies.com/us/en/home/references/protocols/cell-culture/stem-cell-protocols/ipsc-protocols/feeder-dependent-culture-hescs.html Cell number], [http://www.lifetechnologies.com/us/en/home/references/protocols/cell-culture/stem-cell-protocols/ipsc-protocols/feeder-dependent-culture-hescs.html Feeder density] |
Revision as of 00:22, 20 September 2013
- 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.
- Aliquot into cryovials and freeze at –80 °C.
- Refs:Cell number, Feeder density