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Transfection Forum Transfection Forum. Cell transfection discussion including stable cells transfection, transient transfection. Troubleshooting and posting of discussions on lipofection, and reagents: Fugene, lipofectamine, and others.


293T cells death after transfection

293T cells death after transfection - Transfection Forum

293T cells death after transfection - Transfection Forum. Cell transfection discussion including stable cells transfection, transient transfection. Troubleshooting and posting of discussions on lipofection, and reagents: Fugene, lipofectamine, and others.


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Old 09-30-2010, 03:13 PM
Pipette Filler
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Unhappy 293T cells death after transfection



Hi everyone!

I'm new in Molecular Biology and my first task was to do a co-transfection of 293T cells using a LP-IRES2-EGFP vector and a pRSV-Rev and the using Lipofectamine 2000 and Opti Mem with a lot of bad results. First, I had a very poor transfection efficiency until we realize that cells had more than 60 passages, so we change cells and the efficiency get better (around 40%), but my cells start to die, a lot. I change medium after 6 hours of transfection and harvest the cells 24 hours. I also use about 90% of confluency and a DNA - Lipofectamine ratio of 2.5. I'm in crisis. Someone has some idea of whta is happening?

Thanks,
:
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Old 03-20-2012, 08:51 PM
Pipette Filler
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Default Re: 293T cells death after transfection

Hi
No need to use lipofectamine. We been using linear 25 KD polyethyleneimine at 6:1 to DNA for several years now to produce different viruses from transfection 293T with very good results.
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