transfection in MDAMB231 cells - (Sep/19/2006 )
I'm trying to transfect MDAMB231 cells with a plasmid containing GFP with JetPei, but it doesn't work. Does anyone have this cell line? If yes, have you transfected it succesfully? How? Please, help me because I'm quite desperate. I have thought in use lipofectamine or FuGene but I'm not sure they will work, and I don't want to buy them without knowing if they work.
Thanks a lot
We just started using MDA-MB231 too for transfection. From what I have heard from a collegue, they may be hard to transfect. Using Transfast (Promega), he only had about 5% transfection efficiency.
We are then going to try some other transfection with different trial samples: Lipofectamine HTX and 2000 (Invitrogen), Fugene HD (Roche), and Transfast also. I know that it would be possible to get ExGen 500 (Fermentas) too. I tried to ask Qiagen, but they never replied.
I'll let you know if we have good results.
2 other suggestions I recently had: transfecting the cell in suspension after trypsinization and transfecting a linearized plasmid.
I transfect MDA-231s on a weekly basis for luciferase assay and get great results using Lipofectamine 2000. I do experience some toxicity, but the luciferase assay is sensitive enough that I don't worry. Pilot experiments of trasfecting 231s with various reagents and looking for EGFP expression yielded about 40% transfection efficiency. with Lipofectamine 2000. I have not made stables with this line though, only transient assays.