16th May 2019. I have to test a mouse promoter's ability to respond to IL-6 in a mouse renal epithelial cell line. If I transfect normally with Lipofectamine 3000, I have to use Opti-MEM which contains 2% serum and a proprietary mix of cytokines and so these cytokines mask the effect of the IL-6 which I add post-transfection. So, I must optimize transfection with Lipofectamine 3000 using 0% serum or 0.1% serum . . . this will allow addition of IL-6 post-transfection and effect of IL-6 can be dissected. I want to ask if someone has done Lipofectamine transfection without serum . . . do the cells survive this trauma or is the Lipofectamine too toxic in the absence of serum?
It depends how long you keep Lipofectamine, how dense your cells are, and the amount of (volume) cell culture medium etc.
Can try with Ham's F12 nutrient mixture containing N2 supplement. I used this mixture as SFM for 48 hours for primary cells isolated from rat mesenteric artery but not while transfection and did not test the effect of any GFs and or cytokines.
Article Adventitial Neuronal Somata
- Somasundaram C etal. 2006.
Lipofectamine's ability for transfection does not depend on Optimem. You can transfect the cells in serum free medium. The question here is will your cells survive the duration of the transfection without any growth factors in the medium. I generally use DMEM + FBS without antibiotics for transfection with very little optimem but I think an exception should be made in you case.
In osteoblast differentiation by BMP2 for the purpose of their promoter response, I use medium containing 0.2% FBS.
https://www.thermofisher.com/us/en/home/references/gibco-cell-culture-basics/transfection-basics/factors-influencing-transfection-efficiency.html
Hi,
You can use Lipofectamine 3000 in serum-free medium as well...I have performed some preliminary experiments with 'serum and antibiotic'-free alpha MEM (for transfecting human primary monocytes and mesenchymal stem cells). It works fine! You can definitely proceed without using Opti-MEM without a doubt. You need to optimize the protocol to achieve the delicate balance of 'incubation time with lipofectamine' and 'lipofectamine conc.' during transfection. It is doable though.
All the best!
Thanks, Shubhangi Gavali , I have tried it without serum and it does work . . . For my cells (IMCD-3 mouse renal epithelial cell line), 4 hours transfection duration in 24-well plate wells with 25,000 to 30,000 cells per well and 2uL lipofectamine and 1ug plasmid DNA in a total volume of 100uL worked fine. Thanks for your help :)