Culturing Hep G2 Cells - (Sep/27/2006 )

Hi I need help and advice on culturing Hep G2 cells.

The culture medium that I am currently using is Invitrogen's Dulbecco's Modified Eagle Medium (DMEM) (1X), liquid (4.5 g/L D-glucose) Contains 4,500 mg/L D-glucose and sodium pyruvate. Without L-glutamine.

The problem that I am facing is the cells that I got will not attach and those few that attached will have very slow cell growth (almost negligible).

I got the cells from another lab that is using low glucose DMEM instead of the high glucose that I am using now. Currently I am trying to use the low glucose DMEM and gradually change to the medium that I am using now (the reason is I am culturing Hep G215 as well and they are growing fine in high glucose DMEM).

So do you think the glucose contents is the cause of my problem?

Thanks for any help or advice given.

---

do you use serum?
I use a DMEM without sodium pyruvae and thats goes well with same Glucose concentration as yours.

---

Cell density and how well did you disperse cells are very important for this line. Try to culture them with a high cell density to start with. Tryspinize it 37 C 5 min, vortex or pipeting till cells separate from each other.

---

The glucose shouldn't be a problem for the HepG2. I'm using pretty much the same media as you are (with FBS) for my HepG2 cells. From experience though, it seems that you may need to subculture your HepG2 at a higher density. I find I usually need to subculture it into a slightly higher density than some other cell lines such as 4T1 and NIH-3T3 for optimum growth. ATCC recommends subculturing at a ratio between 1:4 to 1:6.
Hope this helps.

---

Hi,
I use Irvine Scientific RPMI 1640 medium with L-glutamine + %10 fetal bovine serum. They grow very well with this formulation. My main problem on this cell line, there is some bubbles (maybe lipid granules) occurs in cell sitoplasm after 3 weeks. Do you have any idea what they are? thanx
Görkem

---