I did qRt-PCR for rat MSCs which are cultured on either growth (GM) or chondrogenic medium (CM). Expression levels of gene of interest (SOX9, Aggrecan, gene markers for chondrogenic differentiation) are higher in GM cultured cells, rather than CM cultured cells. Is there anyone who observed similar pattern with any of differentiation medium?
Hi Gunel
Its strange to know undifferentiated cells express more chondro markers than differentiated one.
Surprisingly there s paper showed (https://www.karger.com/Article/FullText/350089) rat MSCs have high sox9 expression. (It s well know fact that collagen x expression s high in undifferentiated MSCs but interesting to know high sox9 and other chondro markers).
Might be your chondro diff media induce hypertrophy. this leads to down regulation of chondrogenic markers. Just check the hypertrophy markers as well.
COL X s suppose to be high in undifferentiated cells and should go down during initial days of differentiation (like 1st week)
Could be please elaborate your protocol like media used etc..
Hi Rajagopal,
Thanks for the answer, I will consider your suggestions.
hi ozum,
it may be useful to check marker expression not only at the endpoint of your experiment (usually after 3 weeks) but also at some earlier time Points (3d, 7 d, 14d for example). often, cells downregulate mRNA at termianl differentiation. It is more work, but provides valuable information.
best,
Herbert
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