I would like to mix the differentiation-medium (adipogenesis/osteogenesis) for murine MSC myself. I found several publications about this, basically it comes down to:
Basal medium + FCS + antibiotic +
Adipo: 5 μg/ml insulin, 50 μM indomethacin, 1 μM dexamethasone, 0.5 μM 3-isobutyl-1-methylxanthine;
Osteo: 2 mM L-glutamine, 20 mM β-glycerol phosphate, 50 ng/ml thyroxine, 1 nM dexamethasone, 0.5 μM ascorbate 2-phosphate
In 3 publications Sigma was mentioned as producer. I talked to them and got told, that most of the substances are not available in cell culture standards, so I would have to try it at my own risk.
What would you do? Try it? Can you recommend another company? Any suggestions or corrections about the protocol?
Thank you!
Gibco company provides differntiation media (ost/chondro/adipo)-may be tried, we get goog results in canine and rabbit MSCs..
Thank you, already used Gibco, for several reasons I want to mix the medium myself.
only main difference between Ost and Chond differentiation media is FCS which is absent in latter.
Dear Mathias
Regarding the elements for osteogenic diff, Sigma has at least the dex and beta-GP suitable for cell culture. You can try the ascorbic acid salt from WAKO. I think these three elements are the essential for robust osteogenic diff using Dex.
In what concerns adipogenic diff, I don't have that much experience so I can't really comment on the protocol.
Hi, I made up the MSC differentiation media on my own, as you need to do, by using the reagents from Sigma. I got very high efficiency of osteogenic, adipogenic and chondrogenic differentiation of MSC form different sources, ranging from mouse, human and rabbit, therefore I can totally recommend them. Please check out this for further info http://www.ncbi.nlm.nih.gov/pubmed/19298168 and do not hesitate to contact me if you need advice on the protocols.
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