A-431,HepG2- someone working with them? - (Jan/27/2007 )

hi friend,
i am very new in cell culture.presently i am handling A431 SKIN CELL LINE, Hep g2 hepatoma cell line.but i don't know what is the doubling time of a431 & Hep g2 cell line?plz tell me the answer of this basic question which i am asking to all of you.


thanking you. Ritesh k shukla

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From my experience, HepG2 cells doubling time is variable base on confluency. It would take forever to grow them when they are sparsely populated, I would say near 48hrs doubling time. But once they become 30-40% confluence, they will fill the flask overnight. Also, HepG2 cells metabolize very fast, you will need to change your media on a daily basis. Even if you still see pink media from previous day, if you don't change it, you may regret the next day when your media become yellow (this is not related to contamination).

HepG2 cells are also tough attaching cells. They will only come off the plate after 15 min of trypsinization. You can not over trypsinize these cells. It is even harder to break them apart into single cell suspension than it is to lift them off the plate. You will need to pippet them vigorously to break them apart, if you don't, they will grow in clumps. I once forgot to leave them in trypsin for 4-5hrs and they still seeded just fine. Make sure you have enough trypsin to cover the whole surface (1ml for T25, 2ml for T75 flasks), you don't want to have to add more trypsin to the flask because the middle of the flask did not get enough trypsin.

I don't know anything about A431 cells.

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