3 replies to this topic

[lm_choo]

hi,

Recently I am doing transfection on liver cancer cells via Lipofectamine reagent. Since liver cancer cells is not easy to do transfection, may I know is there anyone succesful transfect liver cancer cells via lipofectamine and with what condition?

I understand that lipofectamine will be toxic to cell and I did play again with the concentration of lipofectamine and concentration of DNA in transfection. But it din show a very good result, may I get some consult?

THank you very much
good day

[bob1]

In my experience Lipofectamine is quite toxic to all the cell types I have transfected (I don't think I have done any liver ones though), but Fugene is much less toxic, if that's any help.

[lm_choo]

bob1, on Nov 22 2009, 03:39 PM, said:

In my experience Lipofectamine is quite toxic to all the cell types I have transfected (I don't think I have done any liver ones though), but Fugene is much less toxic, if that's any help.

Hi Bob1,

Thanks for your reply, currently I only have Lipofectamine to try on... and another option is Electroporation... Do you have any advice on electroporation method and precaution?

Thanks again...
Good day to you

[gabamaria]

lm_choo, on Nov 21 2009, 09:32 AM, said:

hi,

Recently I am doing transfection on liver cancer cells via Lipofectamine reagent. Since liver cancer cells is not easy to do transfection, may I know is there anyone succesful transfect liver cancer cells via lipofectamine and with what condition?

I understand that lipofectamine will be toxic to cell and I did play again with the concentration of lipofectamine and concentration of DNA in transfection. But it din show a very good result, may I get some consult?

THank you very much
good day

hi Im choo,

actually I was working before with immortalized human hepatocytes (IHH) and i performed always Ca-Phosphate transfection, and it worked pretty well.
i can send you my protocol if you want, it was an optimized one, from nature protocols. I performed this with cDNA expressing GABA receptors, and the cells were fine after transfection. immunofluorescence was perfomed then to see the expression of this receptor and we had nice pictures.

best regards,

Marie