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Open Access Methodology article

Canine muscle cell culture and consecutive patch-clamp measurements - a new approach to characterize muscular diseases in dogs

Henning Christian Schenk14*, Klaus Krampfl24, Wolfgang Baumgärtner34 and Andrea Tipold14

Author affiliations

1 Department of Small Animal Medicine and Surgery, University of Veterinary Medicine, Hannover, Germany

2 Department of Neurology, School of Medicine, Hannover, Germany

3 Institute of Pathology, University of Veterinary Medicine, Hannover, Germany

4 Centre for Systems Neuroscience (ZSN), Hannover, Germany

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Citation and License

BMC Veterinary Research 2012, 8:227  doi:10.1186/1746-6148-8-227

Published: 21 November 2012

Abstract

Background

The recognition of functional muscular disorders, (e.g. channelopathies like Myotonia) is rising in veterinary neurology. Morphologic (e.g. histology) and even genetic based studies in these diseases are not able to elucidate the functional pathomechanism. As there is a deficit of knowledge and skills considering this special task, the aim of the current pilot study was to develop a canine muscle cell culture system derived from muscle biopsies of healthy client-owned dogs, which allows sampling of the biopsies under working conditions in the daily veterinary practise.

Results

Muscular biopsies from 16 dogs of different age and breed were taken during standard surgical procedures and were stored for one to three days at 4°C in a transport medium in order to simulate shipping conditions. Afterwards biopsies were professionally processed, including harvesting of satellite cells, inducing their proliferation, differentiating them into myotubes and recultivating myotubes after long-term storage in liquid nitrogen. Myogenic origin of cultured cells was determined by immunofluorescence, immunohistology and by their typical morphology after inducing differentiation. Subsequent to the differentiation into myotubes feasibility of patch-clamp recordings of voltage gated ion channels was successfully.

Conclusion

We have developed a canine muscle cell culture system, which allows sampling of biopsies from young and old dogs of different breeds under practical conditions. Patch clamp measurements can be carried out with the cultured myotubes demonstrating potential of these cells as source for functional research.

Keywords:
Myotubes; Voltage gated ion channels; Functional; Dog; Animal models