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An alternate protocol for establishment of primary caprine fetal myoblast cell culture: an in vitro model for muscle growth study

An Erratum to this article was published on 03 July 2013

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.

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Acknowledgments

This work was carried out under the project supported by Competitive Research Grant (C2132) under National Agriculture Innovation Project (NAIP, Component 4), Indian Council of Agricultural Research (ICAR), New Delhi. The financial help in the form of Institute Scholarship to SPS during his Ph.D. study is also acknowledged. Authors are grateful to Vishakh Walia, Senior Research Fellow, Genome Analysis Lab, AG division, IVRI, Izatnagar, Bareilly, India for critical reading of the manuscript.

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Correspondence to Abhijit Mitra.

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Satyendra Pal Singh, Rohit Kumar, and Priya Kumari contributed equally to this work.

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Singh, S.P., Kumar, R., Kumari, P. et al. An alternate protocol for establishment of primary caprine fetal myoblast cell culture: an in vitro model for muscle growth study. In Vitro Cell.Dev.Biol.-Animal 49, 589–597 (2013). https://doi.org/10.1007/s11626-013-9642-0

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Keywords

  • Fetal myoblast
  • Cell culture
  • Desmin staining
  • Goat
  • Myostatin
  • Myogenic regulatory factors
  • Gene expression