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hep-2 & Hep-G2 cell line


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18 replies to this topic

#16 luminb

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Posted 18 February 2005 - 11:21 AM

by the way, in terms of transfection efficiency, hepG2 cells are not very easy to work with. I used double amount of DNA for transfection of hepG2 than that of 293 cells. for example, 1 ug DNA for each well in a 12 well plate. Use more lipofectamine than 293 cells. fugene is fine too.

#17 humptidumpti

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Posted 05 March 2005 - 02:01 AM

Hi everybody,
I am working with HepG2 cells, I grow them in RPMI1640 with 25mM L-glutamin, 10%FCS and 1%P/S. They are slow growing but they form more or less nice layers if you trypsinize them often and long enough. I trypsinize them for 15 min at 37°C in the incubator without touching the fleak to avoid clump formation. But even after 15min cells can stick to each other but this can be overcome by pipetting them up and down..
I am transfecting HepG2 via elcetroporation wich works quite well (up to 40% efficiency)..
paul

#18 Joana

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Posted 29 March 2005 - 12:13 AM

Hi!

May I ask your optimization for transfection of HepG2 via electroporation (cell density, electroporation controls etc.)? I have never tried it but I think it sounds really promising!

Thanks in advance!

Joana

Hi everybody,
I am working with HepG2 cells, I grow them in RPMI1640 with 25mM L-glutamin, 10%FCS and 1%P/S. They are slow growing but they form more or less nice layers if you trypsinize them often and long enough. I trypsinize them for 15 min at 37°C  in the incubator without touching the fleak to avoid clump formation. But even after 15min cells can stick to each other but this can be overcome by pipetting them up and down..
I am transfecting HepG2 via elcetroporation wich works quite well (up to 40% efficiency)..
paul

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#19 Joana

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Posted 29 March 2005 - 12:17 AM

Hi Stefan!

My transfection efficiency has also been under 5%. I have heard that it could be somewhere near 20-30% but never managed to get that myself. B) I havenŽt tried Effectene but IŽll try oligofectamine (invitrogen) soon. IŽm also interested in electroporation. Have you tried it?

With best, Joana

Thanks stefan. Maybe I should also try to grow them without antibiotics.
I have transfected HepG2 with Fugene6 (Roche), but  not very successfully. I think they are in general very difficult to transfect (20-30% of cells) or so I have heard.

If anyone has optimized transfection to HepG2 cells, please let me know too!

Thanks! :blink:

Hi Joana,

this week I also tried to use Fugene6, but the effeciency was below 5%.
May I ask you for your optimization?

Have you ever tried Effectene by Qiagen? This sounds promising and therefore I wanted to test next.

Bye,
Stefan

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