Hi,
I am looking for a technique to label the MSCs mineralization foci in vitro. As I use a Ca/P-containing substrate, I can't use Alizarin Red o van Kossa staining... I have found that OTC has been administered in vivo to label new bone formation.
Does anybody have any experience with the use of Oxytetracycline in vitro to mark MSCs mineralization ?
Thanks,
Laura
That's a pretty cool idea. How would you detect the OTC though? It doesn't have a color. You could dissolve and quantify on HPLC, but that doesn't seem worth while. Neither is using 3H labelled OTC.
I guess your best bet would be fluorescence. That has been done before, even in vivo, but you might want to pick a different tetracycline. Actually regular tetracycline might not be bad:
http://onlinelibrary.wiley.com/enhanced/doi/10.1111/j.1469-7580.2010.01237.x/
Would be cool if this works, keep us updated if you try something :-)
Hi! thanks for your reply and for the paper.
My idea was to visualize it in fluorescence... However i have just realized that it is not a quantitative method... :( I might quantify the fluorescent area but it is not very accurate... so perhaps it doesn't suit my study.
Any other idea or suggestion...?
Laura
Hi Laura, you could probably do this quantitatively, just do the assay in well plate and measure it with a fluorescent plate reader (if the new bone is on top). If your Ca/P phosphate gives too much interference or your construct is not 'well-shaped' you might just dissolve everything with an acid and/or EDTA and then measure it in a plate or cuvette reader. Or do HPLC if you have access to one.
I still think it's worth a try. I'm not in bone-tissue engineering myself, so I don't really know what people normally use. Maybe you can get a method-paper out of it too :-)
Hi Hendrix! Thanks for your help! Definitively I will try and I will keep you upadated :)
Laura
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