I have transfected HEK293 cells to express Protein-GFP conjugate, transfection rate was high (>90%), but the cell morphology changed, many of the cells appear round, but not floating, and they are fluorescent (GFP filter). The cells were transfected using CaCl2+BBS, two days ago.
So, is the morphology change normal?
Thank you.
Need further conditions about the cell confluency and how much plasmid transfected.
No more than 10ug for the 100-mm plate, and calibrate the respective plasmid according to the ratio of their diameter for different size of the 6/12/24 well plate you used.
Be careful about the endotoxin that might be existing in your plasmid aliquot.
I'd like to refresh the medium 6-8 h post Ca2+ transfection to relieve the toxicity.
Generally, 293T become round when they are feeling 'unhappy', try to make them happy then.
I used 10ug for 50 mm plate, about 40% confluency. The cells are regular HEK293, not 293T, thanks!
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