I have a problem with the staining of my cells. I am isolating intestinal myofibroblasts (iMF) from mouse large and small bowel and to address the number of these cells after isolation and during cultivation, after passaging, I perform flow cytometry analysis. I use anti-aSMA, anti-vimentin and anti-CD90 to detect iMF.
aSMA and vimentin stain nicely, but anti-CD90 does not work properly. It has been reported that iMF are positive for CD90, FACS blots in several publications show more than 80-90% of aSMA+ iMF-like cells being CD90 positive. Now in my case I see only 20-25% of aSMA+ iMF-like cells being positive for CD90.
I tested the antibody on thymoid cells (isolated from mouse thymus) and it stains nicely (as expected). Accutase treatment, medium, collagenase and dispase, used for passaging, cultivation and in isolation from the IG lamina propria respectively, have no effet on CD90 signal on the thymoid cells.
Has anyone an idea what the reason for this discrepancy might be? Why does the anti-CD90 antibody not work with iMF, but thymoid cells show expected results?
Thank you very much for a feedback!
Edited by Myofibroblast, 24 March 2014 - 10:05 AM.