mesenchymal stem cell differentiation into osteoblasts (alizarin) - (Nov/06/2008 )
Hi,
We culture mesenchymal stem cells and deal with their differentiation into osteoblasts. For this, we supplement the media with beta glycerophosphate, ascorbic acid and dexamethazone. When I tried to do the alizarin red staining after 28 days, it did not work, thus showing that there was no calcium deposition- making me believe that there was no differentiation of the stem cells. Has anyone encountered such a problem? My beta-glycerphosphate is the disodium pentahydrate salt.... could that be a problem? What other reasons could there be? My alizarin was ordered in solution form and is ph4.2 and is 2%. Any inputs will be welcome. thanks
-melatonin-
QUOTE (melatonin @ Nov 6 2008, 04:50 PM)
Hi,
We culture mesenchymal stem cells and deal with their differentiation into osteoblasts. For this, we supplement the media with beta glycerophosphate, ascorbic acid and dexamethazone. When I tried to do the alizarin red staining after 28 days, it did not work, thus showing that there was no calcium deposition- making me believe that there was no differentiation of the stem cells. Has anyone encountered such a problem? My beta-glycerphosphate is the disodium pentahydrate salt.... could that be a problem? What other reasons could there be? My alizarin was ordered in solution form and is ph4.2 and is 2%. Any inputs will be welcome. thanks
We culture mesenchymal stem cells and deal with their differentiation into osteoblasts. For this, we supplement the media with beta glycerophosphate, ascorbic acid and dexamethazone. When I tried to do the alizarin red staining after 28 days, it did not work, thus showing that there was no calcium deposition- making me believe that there was no differentiation of the stem cells. Has anyone encountered such a problem? My beta-glycerphosphate is the disodium pentahydrate salt.... could that be a problem? What other reasons could there be? My alizarin was ordered in solution form and is ph4.2 and is 2%. Any inputs will be welcome. thanks
Did your cells look different after 28 days? I usually see even without the staining that my differentiation worked.
Where do you isolate your MSCs from? Did you differentiate them into other lineages as well? Did the others work well?
What concentrations of the supplements do you use?
Did you check your stainig with some positive controls?
-vista-
i need to know the difference between Van Kossa and Alizarin Red staining
-E-coli-