Myogenic Differentiation Potential of Mesenchymal Stem Cells Derived from Fetal Bovine Bone Marrow

Abstract

The myogenic potential of bovine fetal MSC (bfMSC) derived from bone marrow (BM) remains unknown; despite its potential application for the study of myogenesis and its implications for livestock production. In the present study, three protocols for in vitro myogenic differentiation of bfMSC based on the use of DNA methyltransferase inhibitor 5-Aza-2'-deoxycytidine (5-Aza), myoblast-secreted factor Galectin-1 (Gal-1), and myoblast culture medium SkGM-2 BulletKit were used. Plastic-adherent bfMSC were isolated from fetal BM collected from abattoir-derived fetuses. Post-thaw viability analyses detected 85.6% bfMSC negative for propidium iodine (PI). Levels of muscle regulatory factors (MRF) MYF5, MYF6, MYOD, and DES mRNA were higher (P < 0.05) in bfMSC cultured under 100 µM of 5-Aza compared to 1 and 10 µM. Treatment of bfMSC with 10 µM of 5-Aza resulted in down-regulation of MYOD mRNA (Days 7 to 21) and up-regulation of MYF6 (Day 7), MYF5, and DES mRNA (Day 21). Gal-1 and SkGM-2 BulletKit induced sequential down-regulation of early MRF (MYF5) and up-regulation of intermediate (MYOD) and late MRF (DES) mRNA. Moreover, DES and MYF5 were immunodetected in differentiated bfMSC. In conclusion, protocols evaluated in bfMSC induced progress into myogenic differentiation until certain extent evidenced by changes in MRF gene expression.

Myogenic differentiation of primary myoblasts and mesenchymal stromal cells under serum-free conditions on PCL-collagen I-nanoscaffolds

“…Unlike Mb, mesenchymal stromal cells (MSC) can be expanded widely without losing their differentiation ability. MSC are known to secrete several growth factors involved in the muscle regeneration process and to stimulate myoblast migration, proliferation, and cell survival [ 9 ].…”

Tissue Engineering for Clean Meat Production

“…A recent report described a first stable culture of bovine embryonic stem cells (ESC) (Bogliotti et al, 2018;Yuan, 2018), which could potentially differentiate into all cell types required to recapitulate skeletal muscle development (Du et al, 2010a;Thorsteinsdóttir et al, 2011;Yan et al, 2013;Chal and Pourquié, 2017), and form a cell bank with an unaltered and stable karyotype, eliminating further dependence on animals for cell isolation. Mesenchymal stem cells (MSC) are another promising candidate, due to their abundance and roles during muscle development (Du et al, 2010b), and their ability to differentiate into myocytes, adipocytes, fibroblasts and ECs (Oswald et al, 2004;Bosnakovski et al, 2005;Huang et al, 2012;Yan et al, 2013;Jana et al, 2016;Ramírez-Espinosa et al, 2016;Okamura et al, 2018). During muscle development, the majority of MSCs commit to the myogenic lineage, generating muscle fibers and a pool of satellite cells.…”

The effect of extracellular matrix on myogenic induction of adipose mesenchymal stem cells

“…ADSCs after ECM ampli cation were inoculated in cell bottles with a density of 15,000 /cm 2 and labeled as group A, while the same generation of ADSCs without ECM ampli cation were inoculated in cell bottles with the same density and labeled as group B and group C.On the second day of cell inoculation, A medium containing 10umol/L 5-azocytosine was added to group A and group B to induce cell myogenic differentiation [11], and the same dose of ordinary medium without 5-azocytosine was added to group C. After three days, cells were cultured in normal medium without 5-azocytosine, and the liquid was changed every two days until the cells were full of trypsin for digestion.…”