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

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BMC Veterinary Research

, 8:227

Cardiovascular, gastrointestinal, urological medicine


BackgroundThe 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.

ResultsMuscular 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.

ConclusionWe 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.

KeywordsMyotubes Voltage gated ion channels Functional Dog Animal models AbbreviationsAqua destDistilled water

°CDegree Celsius

CaClCalcium Chloride

Cat. No.Category number

CO2Carbondioxide, cm, Cubic-centimeter

DMSODimethyl sulfoxide

idIdentification of biopsy donor


DABCO1, 4-Diazabicyclo 2.2.2 octane

DAPI4′, 6-diamidino-2- 327 phenylindole, dihydrochloride

DMEMDulbecco’s modified Eagle Medium

FBSFoetal Bovine Serum


HEPES3.6 g N- 2-hydroxyethyle piperazine-N-2- ethansulphonic acid


KClPotassium Chloride

KOHPotassium hydroxide


MgCl2Magnesium Chloride

MΩMega Ohm







NaClSodium Chloride

NaOHSodium hydroxide

PSGPorcine Skin Gelatine, PFA, Paraformaldehyde

PBSPhosphate Buffered Saline, RT, Room Temperature

TBSTRIS-buffered saline.

Electronic supplementary materialThe online version of this article doi:10.1186-1746-6148-8-227 contains supplementary material, which is available to authorized users.

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Autor: Henning Christian Schenk - Klaus Krampfl - Wolfgang Baumgärtner - Andrea Tipold


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