I would like to immortalise primary MEFs. I have conflicting approaches for how best to do this. First is to take the MEFs through oxidative stress by incubating at atmospheric oxygen levels. However, I feel that this approach is less controlled as we do not know the specific mutations occurring to cause immortalisation. Second is to transfect with SV40 large T antigen. This approach is more controlled in the sense that I know exactly what is causing the immortalisation, however there could be off target effects of this that could affect cell biological processes important to me (I am investigating macropinocytosis). What are peoples thoughts? Would anyone know if SV40 large T affects macropinocytosis?
Alternatively, if anyone knows of a different method for immortalisation I would be very keen to hear. Thanks!
Answer sourced from elsewhere. In case anyone is interested, based on previous publications, I opted for the SV40 large T transfection route.
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