Transfection of human skin fibroblasts - (Jun/19/2006 )

Hi,

We have been trying to transiently transfect human skin fibroblasts. We have tried different transfection reagents like FuGENE 6, Lipofectamine 2000, SiPORT NeoFX, and QIAGEN's Effectene and Super Fect transfection regeants without success.

Does anyone have experience with transfection of fibroblasts? Any advise or suggestions are appreciated.

Kat

(i dont have experience with fibroblasts). As a suggestion, U could try calcium phosphate transfection, if u havnt tried.

Hi,

Thanks for the suggestion. I will try it see.

Kat

QUOTE (katanga @ Jun 19 2006, 03:33 PM)

Hi,

We have been trying to transiently transfect human skin fibroblasts. We have tried different transfection reagents like FuGENE 6, Lipofectamine 2000, SiPORT NeoFX, and QIAGEN's Effectene and Super Fect transfection regeants without success.

Does anyone have experience with transfection of fibroblasts? Any advise or suggestions are appreciated.

Kat

I recall years ago I did that on foreskin fibroblast with Geneporter with some sucess using luciferase as reporter. The number for luciferase activity was decent. I dont know if you want to give a try.

I only transfected mouse fibroblasts and I managed with Lipofectamine+Plus

fibroblasts... uff... that looks difficult.
I had one ore two transfectant per 10 cm plate using fugene hahah
as dnafactory said, maybe lipofectamine works better
anyway, i changed to retroviral transduction with great difference in results

I got 10-E8 total luciferase counts for a 48-well using pcDNA3Luc.

Correction. 10-E7 per well instead.

sorry, i didnt get it
is 10-E7 = 10000000?in a 48 well? or is it 1/10000000?
with what reagent?