SHSY5Y cell transfection - (Sep/04/2006 )

Hi

I would like to transiently transfect SHSY5Y cells. I know they are not very easy transfect and I read one article where they used Lipofectamine 2000 but on invitrogen protocol pages I did not see any information for SHSY5Y cell transfection. anybody has an experience? and maybe protocol?

thanks in advance

Burcu

Hi
I regularly transfect these cells using Fugene6, it is a very simple protocol that can be found on the web site of Roche. Good luck

Look for protocol for transfection of other cell types in invitrogen's site like 293's and follow it.

good Luck !!!

I used TransFast from Promega for different types of cells (including neuroblastoma). Although its procedure is not easy as FuGene or Lipofectamine, it works quite well. It is not so much expensive compared to other reagents. You can try...


Good Luck

I use SHSY5Y cells frequently. They're a bit tricky. I've used a number of transfection reagents, (fugene6, fugeneHD, lipofectamine2000, neurporter, neurofect, turbofectin) and fugeneHD and lipofectamine2000 have worked well. The smallest ratio worked just fine when I tried it, but some constructs get transfected better than others, just keep in mind that the more reagent you use, the more toxic it is to the cells. Also, Fugene HD might not work in serum-containing medium, I transfect in OPTI-MEM for 4-6hr before switching to antibiotic containing-serum-containing medium. Just follow the protocol for the size of your plates, but before transfecting split the shsy5y into plates coated with fibronectin at 1:100, it seems to help them grow and mature better. Transfect at 80-90% with FugeneHD or at 75% with Lipofectamine. Also keep an eye on how fast they grow because they grow quicker with fibronectin, so the transfection complex should be left on for more than 48hrs. These things have helped me a lot. Also, don't use the cells past their 20th passage, or so I've read and been warned. They start loosing their neuronal characteristics.

I mean to write that the transfection complex shouldn't be left on for more than 48hrs (with lipofectamine), or you can just leave it on for 6 hrs and then change the medium. The protocol might explain it much better than I can. They offer several options.