Hello everyone.
I will have to generate MEF (mouse embryonic fibroblasts) from mice of two different genotypes, and to make them become immortalized following the protocole 3T3. The goal is to study the kinetics of the growth of the MEF through the whole immortalization process and to compare the 2 genotypes for this kinetics.
Currently I have a "genotype 1" female mouse of 8 months old and a "genotype 2" female mouse of 2 months; males of each genotypes are more or less 6 months old. I read somewhere that the best age for mating is between 3 and 6 months, so males are OK, but for the females... the question is :
Do you think I can compare the MEF I will obtain for genotype 1 and 2 using these mice or would it be wiser to wait to obtain female mice with more or less similar birthdate ?
Best
MEFs are derived from embryos at about E14, so the age of the mothers should not matter. As long as the mice get pregnant, you should be fine. We usually mate mice by 8 weeks, females can be even sooner 5-6 weeks. One thing though, if the males and females get old and have never been mated , they may not be very sexually active.
