cytoplasmic vacuoles in HepG2 - help with cytoplasmic vacuoles in HepG2 (Apr/04/2005 )

Recently our lab have acquired HepG2 cells from ATCC. I have propagated the cells as descrived by ATCC. But I am worried since this cells show many cytoplasmic vacuoles. I have red that it can be due to several reasons. So I would like to know if someone could help me with this problem. Does somebody know what the reason may be? Is it normal in early passes?

Thanks in advance.

Does your cell look dark?

Dark? I don't understand what do you mean. When they are forming clusters they can look a little darker due to the density of cells.

one of my cell lines also has a few vacuoles. and i read a few papers about the vacuoles and it seems that they result from the toxins generated by some microorgnisms, and there are more vacuoles when the cell density is too low. that's all what i know and i don't know if they have any adverse effect on cell growth.

hi
did you check about mycoplasmas contamination? (it's just a PCR procedure using culture supernatant and give quick reply). That would maybe explain the vacuoles.
I don't have vacuoles in "my" HepG2 cells

Thanks everybody.

I have asked to ATCC. Their reply is that it is not unusual to observe vacuoles in this cell line, as these cells show secretion vacuoles.

In other sources I have red that vacuoles can be due to nutrient defficiencies, etc...

I will try the mycoplasma test and also to use a new-made medium. I will explain what the result is.

Hi

The problem seems to be some kind of nutrient defficiency. I made a new medium, using new supplements (not the old aliquoted and stored ones), this is DMEM, L-Gln, P/S, FBS, NEAA, and the large vacuoles have disappeared by now. It seems that hepG2 show this kind of vacuoles in the growing phase, but once arrived to confluency, the cellls seems to be really nice, with little clumping and small vacuoles, which is normal as these are secretory vacuoles.

Hope I have help.