Despite its potential use in the research of myogenesis and its implications for livestock production, the myogenic capacity of bovine foetal MSC (bfMSC) generated from bone marrow (BM) is unclear. The DNA methyltransferase inhibitor 5-Aza-2′-deoxycytidine (5-Aza), the myoblast-secreted factor Galectin-1 (Gal-1), and the myoblast culture medium SkGM-2 BulletKit were utilised in the three in vitro myogenic differentiation methods employed in the current investigation. Foetal BM obtained from foetuses generated from abattoirs included plastic-adherent bfMSC. Propidium iodine (PI)-negative bfMSC were found in 85.6% of samples during post-thaw viability tests. MYF5, MYF6, MYOD, and DES mRNA levels were greater (P < 0.05) in bfMSC grown under 100 M of 5-Aza compared to 1 and 10 M of 5-Aza. Treatment of bfMSC with 10 M of 5-Aza led to the up-regulation of MYF6 (Day 7), MYF5, and DES mRNA (Day 21) and the down-regulation of MYOD mRNA (Days 7 to 21). SkGM-2 and Gal-1 Early MRF (MYF5) was sequentially downregulated by BulletKit, whereas intermediate (MYOD) and late MRF (DES) mRNA were sequentially upregulated. Differentiated bfMSC were also MYF5 and DES immunoreactive. As a result of protocols tested in bfMSC, myogenic differentiation progressed to a certain level as shown by alterations in MRF gene expression (Brown et al., 2015).
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