sadly there are very little published protocols for MSC differentiation to tenocyte lineage especially using human derived MSCs. There are a few papers in regards the use of BMP12 for the differentiation (JY Lee et al 2011 and QW Wang et al 2005).
you can try seeding these MSCs on tendon scaffolds and harvest them back to culture and expand. Go for characterization like Decorin, Tenascin C , collagen etc. I am working on same aspect and trials are still going on. regards
I also used DMEM, 10% FBS, 1% L-glut, 1% NEAA and 10mM Ascorbic acid on 70% confluent tenocytes for 24hrs and used tht media to investigte differentiation of both hESC and hMSC. As part of my PhD utilised cocktails of BMP and FGF (Manuscripts currrently being written). In regards chracterisation the Tenascin C, Decorin etc. I also utilised Thrombospondin-4 (THBS-4) with Tenomodulin (TNMD) ( TNMD and THBS-4 based on the Paper by Jelinsky et al 2010 ) along with Tenascin C, Decorin, Collagen 3a1 and Collagen 1a2 as my tenocyte panel. This would indicate presence of gene transcripts I then used TNMD antibodies for immunocytochemistry for protein presence as just because there is gene expression does not always allow protien transcription.
Luckily PhD submitted and all dusted. Good luck with the differentiation.
Thanks, Rekha, for your reply, Best Subhash
Thanks, William, Nice work, any expression you have looked for Scx and Mkx genes, just curious,
Subhash
we did briefly look at SCX (however, most of the literature showed it to be upregulated during mechanical stimulation) therefore we decided to look only at endpoint expression of TNMD, THBS-4 etc. with growth factor induced differentiation we could not detect SCX so it remains a distinct question in regards its role? one hypothesis is it is trancient and our timepoints missed the expression??? but the cocktails we developed worked on both hESC and hMSC (bone marrow aspirate derived)
In addition to the Lee and Wang papers listed above, we also recently published a paper on BMP12 for adipose derived MSC differentiation: PLoS One. 2013 Oct 14;8(10):e77613. The methods and outcomes in that paper may be useful to you.
Thanks, Stavros,
It is interesting paper very clearly showing dose-dependent tenogenic expression of ASCs,
Regards,
Subhash
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