It is my first time working with SH-SY5Y, and I have some questions. I will appreciate any help!
First, I would like to know if low serum (1 - 5%) is able to stop or decrease cell division and, in a transfected cell line, if the DNA can be stable for at least 7 days in this condition (DMEM/F12; 1-5% serum). If it is not possible, for how long the DNA stays stable with high protein expression in a SH-SY5Y line (double time - 48h)?
I ask this because I was wondering if there is another way to make a stable transfection (I just need 7 days of high protein expression) without spending lots of months working on it. I have only 4 months to finish my research and I need to know if there is an easy way to do it.
Second, if I differentiate SH cells with RA 10uM, will they continue to grow or this kind of differentiation stops cell cicle (should I maintain RA until the end of the experiment)?. Can I easily transfect these cells as a second option?
And finally, if there is no way to get rid of the standard protocol, can I use 20% of fetal serum in order to speed up cell growth and get my stable line faster (once sh double population time is 48h)? Right now, I'm using 10% with 600 ug/ul of G418 in DMEM/F12 and I already got some cell colonies (20 days pos transfection in 6 well plates) and now I will try to pick up monoclonal clones.
Thanks in advance!