Hepg2 cells growing on top of each other - (Apr/16/2012 )

I am growing HepG2 cells and in the flask there are islands of what look to be dead cells growing on top of the other cells. They break off in chunks. This has happened a couple of other times in different passages. I am only on passage 8

HepG2 like to gather in clusters when they are left in culture without subculturing for too long. Try to suspend them well during next subculturing to detatch cells from one another by pipetting multiple times. If you are not shure whether you can do it gently enough not to kill the cells while intensively enough to really detatch them, you may divide the original cell suspension into 2 and mix one gently and the other really harsh and then compare the effect.