HepG2 Cell Culture Problem

[Richard Cowart]

I have been culturing HepG2 cells for years. About a 5-6 weeks ago the cell growth rate slowed and the cells showed large amounts of vacuoles within the cytoplasm. Nothing has changed with my set-up, from media to the incubator. I just split a flask that has been growing since 7/11 and there was only about 1/4 of the plate with cells. The CO2 reads 4.9% on the incubator readout. Perhaps I should check this as I have a digital CO2 analyzer. Again, I note that the vacuoles are extremely prominent. Any ideas from anyone?

[celartia]

I don't know the specifics of your protocol but....

....it is a well known fact that culturing cells in hyperoxic conditions (even with 5% CO2) is known to cause a suppression of gene expression & can lead to cell mutation.

So a cell line expanded through several passages in these conditions may cause such issues.

Our Company have patented a product called Petaka which is a plastic disposable cell culture device which auto regulates the oxygen exposed to cells so as to offer physiologically relevant oxygen tension. It is a very unique & innovative technology. We ourselves have had some great HEPG2 cultures over the last few years. would encourage you to check it out.