I gave tried lentiviral infection system but the transfection efficiency is not very good? Kindly suggest me some other methods as well as how can I improve the transfection efficiency in suspension cells using lentiviral system and other methods available ?
Hi Ghulam Abbas, as i know you shouldn't transfect plasmids directly to these AML cell lines, is better to use lentivirus to infect U937 cells, which can get a more than 90% efficiency.
Hi Ghulam Abbas, I agree with the floor above. Lenti-virus is the easiest way to transduce ur cells either primary cell or cell line. If u insist on doing transfection, electroporation is another choice, it could transfect jurkat cells or B cells as high as 30-40% of efficiency, as I know.
I have transfected different plasmids in many hematopoietic cell lines and the more efficient system is the lentiviral or retroviral infection. Maybe you could improve your lentiviral production. I use to recommend tittering your lentiviral particles to adjust the quantity of viral particles to the number of cells you want to transduce (MOI). Sometimes you have to increase the MOI depending on the cell line.
I dont know if you are already using polybrene but it is very useful. Another thing you could do is to concentrate the cells with the virus for 30 min in serum-free media before seeding them to increase the chances of the virus to engage the cells. Good luck.
thank you Ester Molina , Qing Yang and Ezzouhra Elmaaiden for your kind suggestions , I have already tried Lenti-virus method but couldn't get satisfactory results.
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