Transfection of HKC-8 - (Oct/03/2008 )
My lab has been trying to transfect human kidney cells (HKC-8) with PKB. We use lipofectamine 2000 to do this and so far the transfection has only worked once( and failed multiple times). We have been using 1.2 ug of DNA to transfect cells in 6 well plate. We transfect for 5 hours and then remove the media. We used to transfect the cells overnight but we found that this killed most of the cells. When we transfect the cells they are about 70-80 % confluent. Should we increase transfection time or amount of DNA? Should we let the cells become more confluent? IF anyone has any suggestions I would appreciate it! Also we have been contemplating electroporation to trasnfect the cells? If anyone knows where I can find a protocol I would appreciate it. Thanks
I haven't used HKC cells but some suggestions from my experience with other cell lines.
Use lower amounts of lipofectamine. And you could use more DNA for the 6 well plate (we use 2ug per well). Also if there r more cells, it also prevents cell death.