I have two quesitions:
1 I have tested toxicity of Curcumin to MG63 osteosarcoma cells, 6000cells/96well were cultured under 10% FBS medium for 48h. Now I want to change study plan to test the toxicity of 72h or even 96h. However, cell numbers in control well(cells without Curcumin) grow confluent after 72h and inhibit itself.
so can I use 2% FBS? so cells will not confluent to inhibit itself and I still could compare the control group with medicine groups to calculate cytotoxicity.
2 in calculating IC50(LD50), I have to let my inhibition curve to cover 0% and 100% inhibition rate? is 0% to 85% acceptable?
Thanks
Hi Mingjie Wang ,
1. If you are facing problem cell confluence then it may be due to some other factors also. But if you find it due to FBS then still it is not recommended to reduce it by 80% in one shot like 10% to 2% you may try 8% or 5% initially. If problem was due to higher FBS then it will be rectified.
Moreover, try see literature on this how other researchers are using FBS.
2. I think no to your second question because generally it is taken 0 to 100%.
Why you wish to take 0 to 85% ?
Thank you,
Good luck
Hi, Deepak
Thanks for your answer, it is very practical to select a reasonable FBS concentration to have cells not growth too confluence.
For the second question, I found it doesn't make sense know, I performed IC50 test again to have 0-100% inhibition.
Kind regards
Mingjie
1) I do not recommend you reduce FBS because reducing the FBS ratio may change response. Instead you may seed 3000 cells/well.
2) It depends on how you generate your graph but generally yes it is acceptable.
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