I am trying to grow hepatic cell lines like HepG2, HUH-7 and ,Hep-3b for anticancer work.
I am growing all in RPMI-1640 + 10%FBS + 1X Ab/AM. HUH-7, and hep-3b are growing very nicely but HepG2 grows with difficulty.
Any advice?
Because they grow very slowly :). To work with this cell line it is very important to set up very carefully the seeding conditions (for example, 10000 cells/cm2). Change the medium every two days and split them before they reach confluency, since they start to aggregate. I used to culture this cell line in DMEM high glucose, 10%FBS, L-glu, NEAA, and P/S. Be patient and good luck
I use growth media containing DME H-21, 10%FBS and 1%P/S. I split cells 1:3 every two days. It works for me. Good luck.
Thanks Maya vila i follow ur advice. @ seyed i checked for mycoplasma.its negative.
i grow them in dmem with 10%FBS and without P/S. no troubles so far.
For HepG2 is recommended to use DMEM:F12 medium + 10% FBs + 1% antibiotics
be carefull if you are going to use 20% serum, it is not phisiologic (neither 10%, but it is more common) and sometimes could be toxic. Are you sure that your cells are not contaminates...mycoplasm?
Iam changing the media every 2 days and using 10% FBS .but lag phase was more 10days i felt.now growth seems good.thanks to all.
I agree with Agustina, in fact in my experince the micoplasma contamination often affect the cell cycle
I have problems in reaching the confluence.
I seed 50000 cells in the 96 well plate and after 24 h they are not optimally confluent.
I heard someone is leaving the cells for 3/4 days in the 96 well plates before incubate them with the treatment. They seed 10000 cells per well.
Do you think is reliable? any other tips?
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