title: An alternate protocol for establishment of primary caprine fetal myoblast cell culture: an in vitro model for muscle growth study.
abstract: Cultured myoblasts have been used extensively as an in vitro model in understanding the underlying mechanisms of myogenesis. Various protocols for establishing a pure myoblast culture have been reported which involve the use of special procedures like flow cytometry and density gradient centrifugation. In goat, only a few protocols for establishing a myogenic cell culture are available and these protocols use adult muscle tissues which often does not yield sufficient numbers of precursor cells with adequate proliferative capacity. Considering the disadvantages of adult myoblasts, we are proposing an alternate protocol using caprine fetus which does not require any special procedures. In the present study, more than 90-95% fetal-derived cell populations had the typical spindle to polyhedral shape of myoblast cell and stained positive for desmin, hence confirming their myogenic origin. These cells attained the maximum confluency as early as 3-4 d against 3 wk by adult myoblasts indicating a better growth potential. Further, quantitative real-time PCR analysis revealed a higher expression (p < 0.01) of myogenic regulatory factors (i.e., myogenic determination factor 1, myogenic factor 5, and myogenin) and myostatin (MSTN) in the fetal as compared to the adult myoblasts. Consequently, higher proliferation and differentiation ability along with higher abundance of myogenic markers and MSTN make the fetal myoblasts a better in vitro model.
authors.affiliation: Department of Animal Genetics and Breeding, C.o.V.Sc. & A.H., DUVASU, Mathura, 281001, India.
authors.forename: Satyendra Pal
authors.initials: SP
authors.lastname: Singh
authors.forename: Rohit
authors.initials: R
authors.lastname: Kumar
authors.forename: Priya
authors.initials: P
authors.lastname: Kumari
authors.forename: Abhijit
authors.initials: A
authors.lastname: Mitra
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D006041:Goats; D024510:Muscle Development; D032446:Myoblasts / Q000166:cytology* / Q000378:metabolism; D018006:Myogenic Regulatory Factors / Q000378:metabolism; D018008:Myogenin / Q000378:metabolism; D061251:Primary Cell Culture
chemical_list: D018006:Myogenic Regulatory Factors; D018008:Myogenin