Hi, everyone
I have a simple question about timespan of serum starvation and transfection.
Recently I work on U-2OS cells which have doubling time around 20 hours, and before I transfect U-2OS cells, I deprive cells of serum in DMEM for 4-8 hours, depending on what constructs I transfect. For the time being, 8 hours of serum starvation works very well.
I know that the mechanism of raising transfection efficiency by serum starvation is to arrest cells at G0/G1 state, and re-entry of cells into cell cycle boosts cell's intake of nutrients along with transfection reagents and DNA within it.
Now I am preparing to shift from U-2OS to neuroblastoma cells SK-N-AS which has doubling time around 40 hours. This makes me wonder if the timespan of serum starvation should be prolonged (let's say, 16 hours) before transfection.
Thank you
I do transfect cells with serum starvation, that is 0,5% serum in DMEM. I change the medium at the end of the work shift and proceed with transfection the next day morning. I saw no decline in cell condition [primary chondrocytes, fibroblasts and popular cells like MDCK] and I could say 24 hours in this setting would work fine as well. MDCK could survive in fine condition even 48h starvation not to mention HeLa or SiHa.
In my opinion, it is more crucial to adjust the concentration of plasmid correct when you use lipofection.
And, as always, it might or might not work for your cells and must be checked.
Yen-hsun Chang I think overnight starvation is quite sufficient and do work well om most types of cells
Thank you all for offering nice advice. I'll try the conditions this week and see how it works!
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