I did passaging ES cells experiment. When i looked at MEF Feeder Cells (it has been treated by Mitomycin C), it's already mature and it is supposed to be good for ES Cells. However, one day after i use it for passaging ES cells, several MEF cells died and the remaining cells are not good such as the MEF cells become shorter than before ES cells passaging.
Why does MEF feeder cells died after i passage the ES cells? Is it possible if maybe culture medium temperature affect the MEF feeder cells?
May be original feeder not good
or put so much Mitomycin c
also Temperatur effect and also incubation effect
Best regards
What species of ES cell?
What ES cell media are you using?
What duration and concentration of Mitomycin C are you using?
What passage number are your MEFs?
What temperature are you using for your cultures?
I forget but now i don't have any problem with feeders. Thank you for your concern
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