I would like to create a fibroblast cell line but I am unsure about some stuff. I want to keep things simple so I plan to obtain MEFs from mouse embryos and transduce them with SV40. Can I use pBABE-puro SV40 LT as my SV40 source? I dont want to buy an immortalization kit since they are crazy expensive.
My MEF isolation protocol uses MEM + supplements for culturing these cells. Could I use DMEM instead? I plan to package the virus with phoenix cells and they are grown in DMEM. I would like to grow both cell lines with the same medium but I dont know if the extra glucose would have a negative effect on the MEFs.
Once immortalized I plan to transduce these MEFs again.
I would appreciate if someone with experience can offer some advice and answer these questions.