Abstract
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.
Highlights
The recognition of functional muscular disorders, is rising in veterinary neurology
Approaches to evaluate functional components of such disorders in cell culture systems have not been established in veterinary neurology so far
Cell culture Biopsies were taken during standard surgical procedures from different striated muscle of 16 dogs, stored for one to three days at 4°C in a special transport medium in order to simulate shipping conditions and were further processed
Summary
The recognition of functional muscular disorders, (e.g. channelopathies like Myotonia) is rising in veterinary neurology. 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. Approaches to evaluate functional components of such disorders in cell culture systems have not been established in veterinary neurology so far. The aim of the current study was to establish an appropriate shipping method in combination with adequate tissue processing for collecting primary satellite cell cultures in dogs. The feasibility of electrophysiological examinations of voltage- gated ion channels expressed by canine myotubes with the patch clamp technique was evaluated
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