I have recovered my stock cells of HEPG2 cells for my experiments and I have noticed it grows slow around 3 days (or 4 days even, if I don't change the medium 48 hrs after passage). It is currently on its 6th passage and I noticed that the morphology of some cells in the flask was starting to exhibit elongated cell morphology (from my experience, these are fibroblast-like characteristics). Although most of the cells (around 95%) still retain HEPG2 morphology.
I use DMEM-Low Glucose supplemented with 10% FBS and 1% antibiotic (penicillin/streptomycin) as my culture medium.
Does anyone have advice or explanation or experience like this? Thank you so much.
Dear Bryan Angelo!
Maybe try using a high glucose medium. since HEP2 cells are cancer cells, and cancer cells are very sensitive to glucose, and it is used as the main energy substrate in the glycolysis process for vital activity.
https://www.nature.com/articles/s41598-019-43669-7
Cell maintenance
The culture of the human hepatocellular carcinoma HepG2 cells from ATCC (HB-8065) was performed in 75 cm2-cell culture flasks in DMEM with high glucose and pyruvate content (Gibco, 41966-029) medium supplemented with 10% fetal bovine serum (Gibco, 10270-106).
I agree with Alexandr Chernov. If there is now reason for using LG medium (like examining matabolism), you should stick with the "original" HG medium.
Good luck!
I did experience it before. It could be due to the medium too acidic and make it alter the morphology. But I'm not 100% confirm. Before we try to troubleshoot for a year, most likely is due to the medium. If I changed the medium 24 hours, and subculture at 70% confluence. Then it will be ok. Unfortunately, if I left the cell for 48hours without changing medium, and subculture at 80%, it will turned to elongated bipolar shape, and at the later passage, it will start to die, and cannot proliferation.
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