i was under the impression that 100% confluent cells grow less, and that around 95-100% confluent cells are ideal for glucose depletion measurement in the media. This is because my hepg2 usually stay at around 50-60% confluent after 1:2 splitting for about a week. I've tried using insulin transferrin selenium supplements and this seems to reduce the doubling time by 1 or 2 days. Unfortunately ive not heard of ITS being used for hepg2.
Can i start measuring the glucose depletion in the cell media (and for future reference flow cytometry) when they are in about 70% confluence? i feel that if i do this the cells may consume glucose due to the log phase which is what i am trying to avoid. thats why i culture my cells to ~100% confluence. I know that the cells may mutate or something, but how extensive can the mutation be if it is only left at 100% for 24hours?
This thesis is being such a pest because my hepg2 grow so slow.