I am growing my HepG2 cells in low-glucose DMEM (1 g/l glucose) with 10% FBS and 1% anti-anti in a T25 flask. The cells were passaged into a T75 flask after they had grown and clustered, as shown in picture #1. However, the growth of the cells is very slow, as you can see in picture #2, which was taken 5 days after passage. Additionally, I recently noticed a shape appearing in the third picture in the flask, but I couldn't discern what it was. Can anyone with experience working with this type of cell explain it to me?
Hi,
Picture #1 is fine in my opinion.
What kind of FBS do you use for your culture? In past, I had problems with specific brands. I use FBS from south America of Gibco or Merck.
How do you detach them? I usually use trypsin 5x and split them between 1 to 3-6 fold.
I do not know what is in picture #3.
Regards
LG
Thank you, Leonardo Gatticchi, for your reply. I used FBS (fetal bovine serum) that was heat-inactivated from Gibco. Is there any concern about using this?
For detaching the cells, I use 0.25% Trypsin-EDTA (1X), but it takes around 10 minutes for the cells to detach.
The shape in picture #3 disappeared. Cross-contamination was suspected in our lab; however, there were no signs of it except for shapes like this one, and many black dots in the background, which I think are debris.
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