iam working on hepg2 cell lines which on passaging incresing in number but not attached to flask and its morphology also little bit changed can you suggest the reasons and tell how to maintain this culture?
I have observed the same problem with HepG2 and they are quiet sensitive cells. Ok, first don't ever allow HepG2 to become overconfluent...Overcrowding of cells result in deadherence and rounding of cells and you will lose all cells after a certain time..so you have to be very active while culturing HepG2....trypsinize them as soon as they reach around 80% confluence...
Second, use alpha-MEM and US grade FBS..they grow well in MEM...CHANGE MEDIA EVERY THIRD DAY without miss