title: 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.
authors.affiliation: a Departamento de Apoio, Produção e Saúde Animal, Faculdade de Medicina Veterinária , Universidade Estadual Paulista "Júlio de Mesquita Filho" , Araçatuba , São Paulo , Brasil.|b Departamento de Fomento de la Producción Animal, Facultad de Ciencias Veterinarias y Pecuarias , Universidad de Chile , Santiago , Chile.
authors.forename: Lucas Hidenori
authors.initials: LH
authors.lastname: Okamura
authors.affiliation: b Departamento de Fomento de la Producción Animal, Facultad de Ciencias Veterinarias y Pecuarias , Universidad de Chile , Santiago , Chile.
authors.forename: Paloma
authors.initials: P
authors.lastname: Cordero
authors.affiliation: b Departamento de Fomento de la Producción Animal, Facultad de Ciencias Veterinarias y Pecuarias , Universidad de Chile , Santiago , Chile.
authors.forename: Jaime
authors.initials: J
authors.lastname: Palomino
authors.affiliation: c Departamento de Ciencias Biológicas, Facultad de Ciencias Veterinarias y Pecuarias , Universidad de Chile , Santiago , Chile.
authors.forename: Victor Hugo
authors.initials: VH
authors.lastname: Parraguez
authors.affiliation: d Departamento de Ciencias Clínicas, Facultad de Ciencias Veterinarias y Pecuarias , Universidad de Chile , Santiago , Chile.
authors.forename: Cristian Gabriel
authors.initials: CG
authors.lastname: Torres
authors.affiliation: b Departamento de Fomento de la Producción Animal, Facultad de Ciencias Veterinarias y Pecuarias , Universidad de Chile , Santiago , Chile.|e Department of Biomedical Sciences and Pathobiology, Virginia-Maryland Regional College of Veterinary Medicine , Virginia Tech , Blacksburg , Virginia , USA.
authors.forename: Oscar Alejandro
authors.initials: OA
authors.lastname: Peralta
mesh_terms: D000818:Animals; D001853:Bone Marrow / Q000196:embryology*; D001854:Bone Marrow Cells / Q000166:cytology* / Q000502:physiology; D002417:Cattle; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D005240:Feasibility Studies; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D024510:Muscle Development / Q000502:physiology*; D032446:Myoblasts / Q000166:cytology* / Q000502:physiology
keywords: Bone marrow; cattle; mesenchymal stem cells; myogenesis