I've been generating and culturing multiple different human iPSCs lines on mito-MEF feeders using standard hiPSC media composed of Advanced DMEM/F12 (Gibco) with 20% KSR, pen/strep, glutamax, 2-mercaptoethanol and daily fresh bFGF supplementation. As advanced dmem/f12 contains non-essential amino acids, I do not further supplement in my culture. So far, so good. I was wondering if anyone has ever tried to reduce the % of KSR in their culture, as Advanced DMEM/F12 already contains in part holo transferrin, insulin, BSA (AlbuMAX II) and most of other essential components that were also provided by KSR itself, and are completely missing in the standard DMEM/F12 basal medium.
Now I am trying to optimize their culture in 15% KSR, but has anyone ever tried even lower concentrations (which would seriously affect the cost of media for routine iPSC culture) of KSR when using Advanced DMEM/F12?
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