Transfecting Human aortic smooth muscles....Help PLEASE - (Dec/28/2005 )
I tried to transfect human orteic smooth muscle cells but unfortunatly I failed. I used different transfecting reagents such as lipofectamine, lipofectaime2000,fugene, and genejuce. All didnt work when I tried to test their effeciency in transfecting HASMCs with B-gal or GFP. Lipofectamin and Lipofectamine2000 were harsh on the cells most of them died.
If anybody succeed in transfecting these cells please tell me in details what exactly i should do?
Similarly, I have used lipofectamione2000, fugene-6, Genejuice, Effectene and JetPEI on my cells and found that these reagents did not work on my cells and caused cell death.
I recently switched to gencarrier-1 on primary aortic smooth muscle and myoblasts and found it is the best one for my cells with ~20% efficiency and low cell loss.
Thanx Bonita for your reply. Exactly from where you got the gencarrier-1 and dont you think th efficiency is too low?.
When you transfect your cells using lipo, and lipo2000, did you dilute the DNA and The transfecting reagent separatly and then incubate them together before transfecting the cells? for how long did you transfect your cells?
waiting for your reply