hMSC from bone marrow how do you expand them before induce adipogenesis?
What types of medium can be used for the expansion? is DMEM-F12 ok?
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Yuri Ciervo
Hi Benedetta, I work with MSC from adipose tissue. I usually start differentiation when they are 90% confluent using DMEM high glucose. First week 1uM dexamethasone, 10uM Insulin, 0.5mM IBMX, 200uMIndomethacin for 4-5 days or until you start seeing differentiation in your culture. Them I continue without IMBX for another 10-12 days. You can use F12 for expansion.
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Shalini Suresh
Hi Benedetta,
I work with tumor derived MSC. I agree with Ciervo, the protocol does work. There are commerically available medias as well from Lonza and Stemcell technologies. I usually start when these cells are 75%-80% confluent.
hMSC from different origins have different time points- hBM-MSC take 18-20 days for differentiation, hAD-MSC differentiate within 12-15 days, hDP-MSC take minimum of 15-17 days and tumor MSC differentaite very quickly within 10 days of induction.
make sure media is replaced every 3 days.
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