I'm using DMEM with glutamine and 10% FBS. Since a month now, the growth rate has seriously dropped. No contamination has been detected. What can I do to make them grow quicker?
You could add 10ng/mL of basic FGF. It might increase growth rate.
The main question is why the growth rate decrease ? If your cells have too many passages it might explain why they do not growth. And it might be difficult to increase growth rate.
When you take about contamination, do you mean that you also have tested the presence of mycoplasma in your culture medium? I also agree with Stéphane about the passage number. In my lab we stop to use fibroblasts cell lines after 10 passages. Concerning the add of other molecule in your culture medium, be carefull with the goal of your study...Adding growth factor or high level of FCS could modify the basal physiology of your cells... Regards, Gwen.
I would strongly suspect mycoplasm contamination. I suggest thawing a new stock of cells--once cells drop in growth rate with no changes in medium, it's time to get out a new batch. Make sure to keep viable stocks of cells in the freezer--if they have been frozen for a long time, take out your stock, grow them up and freeze new stocks every year or two.
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