I'm trying to transfect N1E-115 cells and the HiPerfect protocol advices not to add fetal bovine serum during the transfection phase. Unfortunately, my cells are dying without the FBS. Do you know if I use 2 or 4% in the medium in the HiPerfect mix will it interfere with the transfection?
Serum might influence the formation of the complex between the construct you want to transfect and the lipo-reagent. We use a serum-free medium (such as Optimem) when creating the complex. I prefer to use reverse transfection where you add the comlex to the plate first, and then plate the cells directly on top, using the normal growth medium with serum. Although, some cells are harder to transfect than others, and if your transfection efficiency is low you could try lowering the amount of serum during transfection. Or change the serum-free-medium+transfection mix after 4-6 hours with normal medium.
Hello, very good suggestion from Synnove!
I would simply add that many other reagents are available on the market, and you can find something compatible with FBS serum, like our Viromers.
If you have further questions, do not hesitate to contact me.
https://viromer-transfection.com
Bests, Sandra
Hi Juliana, FBS has a tendency to inhibit the transfection process and it is advisable to use no FBS in your media but there might be a chance to use other supplements which maintain your cells. It is also good to try with lower FBS concentration. good luck.
Hi,
Just to add, I have tried 4%FBS, do not add in mix, suppose if performing 15ml transfection, prepare mix with 7.5ml optimem with your complex forming agents and enhancer and keep at incubation. Another 7.5ml can be another growth media where we can add FBS before hand (remember give sufficient time for cells when resupending it, so preferred do it at least 30 min before hand) then only mix both. Keep at stagnant co2 incubator for 4_5 hrs and then add more 15ml of growth media to avoid toxicity.
As usual incubation in 5%co2.
Hope it would help!
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