Hello I am writing because I am having serious problems transfecting foreskin human fibroblasts with several plasmids at a time. At first we started using the transfection reagent Trans-IT-2020 and the yield was quite good, considering they are fibroblasts ( at least 30-40% cells were transfected).
However, now it is imposible to transfect them and I cannot find the solution for this . We have tried buying a new batch of cells and start with the lowest passages, but still have the same problem.
Has anyone ever had the same problem?
We have had success with the transfection reagent: X-tremeGENE HP (Sigma Aldrich) transfecting mammalian cells.
We investigated several transfection strategies and reagents and X-tremeGENE HP was the most efficient.The XtremeGENE HP transfection reagent is formulated to transfect “hard-to-transfect” cells. An added benefit is that this transfection reagent has very little cytotoxic side effects and can be left on the cells for the entire duration of the transfection experiment - potentially increasing the transfection effectiveness of "hard to transfect" molecules.
Low transfection efficiency in fibroblasts is common - you need special chemical formulations of lipid reagents to get good results (see https://altogen.com/product/fibroblast-transfection-reagent-fibroblast-cells/ ). Several plasmids at a time only strains the cells more, so having a complex condenser at hand should help you improve your results.
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