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Cytotechnology

, Volume 63, Issue 4, pp 399–406 | Cite as

Characterization of in vitro cultured myoblasts isolated from duck (Anas platyrhynchos) embryo

  • He-he Liu
  • Liang Li
  • Xi Chen
  • Wei Cao
  • Rong-ping Zhang
  • Hai-yue Yu
  • Feng Xu
  • Hua He
  • Ji-wen Wang
Original Research

Abstract

Myoblasts isolated from duck embryonic muscle were purified and in vitro cultured. External characteristics were observed by using the immunofluorescence technique, and growth curve of duck embryonic myoblasts was established after measuring with the MTT method. Moreover, mRNA expression of three marker genes, the Desmin, the muscle creatine kinase (Mck) and the troponin C (Tnnc), which could reflect the development status of myofibers, were detected each 24 h for cultured cells by using the qPCR technique. Results showed that the in vitro cultured duck myoblasts went through a series of developmental stages, including the proliferation of myoblasts, the differentiation of multi-nuclei myotubes, and the formation of myofiber. The cultured duck embryonic myoblasts entered into a logarithmic stage approximately on the fourth day after seeding. Accompanying with its progressive growth before entering into the logarithmic phase, the myoblasts also showed some differentiation phenomena, reflected by a low expression level of Desmin and high expression level of the Mck and Tnnc genes. During the rapid growth of the logarithmic phase, there was a high expression of the Desmin gene, and a low expression level of the Mck gene and the Tnnc gene in the cultured myoblasts. The expression profiles of the three marker genes for muscle development could be used for distinguishing the different developmental stages of in vitro cultured myoblasts at the molecular level, which would be more accurate and more feasible than observing the external characteristics of the cultured cells.

Keywords

Gene expression Embryonic myoblasts Cell growth 

Notes

Acknowledgments

The work was supported by the National High Technology Research and Development Program of China (No.2010AA10A109), and Chinese Agriculture Research Service (CARS-43-6).

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • He-he Liu
    • 1
  • Liang Li
    • 1
  • Xi Chen
    • 1
  • Wei Cao
    • 1
  • Rong-ping Zhang
    • 1
  • Hai-yue Yu
    • 1
  • Feng Xu
    • 1
  • Hua He
    • 1
  • Ji-wen Wang
    • 1
  1. 1.Key Lab of Animal Genetic Resources, College of Animal Science and TechnologySichuan Agricultural UniversityYa’anPeople’s Republic of China

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