I have transfected COS 7cells to express Protein-GFP conjugate, transfection rate was very low (20-30 %) , but the cell morphology changed, many of the cells appear round, (nucleus round too) but not floating, and they are fluorescent (GFP filter). The cells were transfected transiently using PEI and incubated for 48 h.
Plasmis:PEI /1:3, different plasmid concentrations, 24 well plate.
while control cells had normal shape (mock control)
So, are these morphological changes normal?
Thank you.
1. did you replace the medium after transfection (~4 hrs)?
2. have tried lower PEI ratio?
3. the gene is toxic?
Dear Junjie
thanks for answering.
1.Yes I changed the medium after 4 hours
2.For pei no , but I tried different pDNA concentrations
3. It is a hypothetical protein (unknown)
I usually use lipofectamine, sometimes, higher lipofectamine kills the cells.
I think PEI may do the same thing.
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