BMSC to osteoblast - (Feb/23/2007 )
Hi all,
Do anybody have protocol of BMSC isolation and differentiation to osteoblast. what are the best markers studied for osteoblast? i had tried dexamethasone treatment on BM mononoclear cells and observed that cells do undergo change in morphology from round to fibroblats like strongly adherent cells. but i had no marker, so couldnyt prove whether those cells were osteoblast or not.
pls help
regards
-poonamd-
QUOTE (poonamd @ Feb 24 2007, 07:49 AM)
Hi all,
Do anybody have protocol of BMSC isolation and differentiation to osteoblast. what are the best markers studied for osteoblast? i had tried dexamethasone treatment on BM mononoclear cells and observed that cells do undergo change in morphology from round to fibroblats like strongly adherent cells. but i had no marker, so couldnyt prove whether those cells were osteoblast or not.
pls help
regards
Do anybody have protocol of BMSC isolation and differentiation to osteoblast. what are the best markers studied for osteoblast? i had tried dexamethasone treatment on BM mononoclear cells and observed that cells do undergo change in morphology from round to fibroblats like strongly adherent cells. but i had no marker, so couldnyt prove whether those cells were osteoblast or not.
pls help
regards
Usually, for osteogenic differentiation of bm-msc, we use also beta-glycerophosphate (10 mM) and ascorbic acid (50 ug/ml) and I think that these are critical for a terminal differentiation that could be reveald by matrix deposition (positive to alizarin-s stain) and by some molecular markers. This reference (http://www.ebmonline.org/cgi/content/full/226/6/507) could help you,it's a minirew about msc and diff potential. I think it's hard to phenotipic characterize this kind of cells, i.e. by cytoflow or IF, I've tried with MoAb agaisnt osteopontin without any convincing results.
Hope been helpfull
-XCyto-