The aim of the present study was to explore a simple and efficient procedure to culture and purify adult canine myoblasts in vitro. Muscle from adult Beagle canines was isolated and harvested by mechanical decomposition and two-step enzyme digestion. The cells were then purified by a method involving differential adherent velocity and flow cytometry. The morphologic properties and growth states of the cells were observed, and the cell phenotype was characterized by flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry. Approximately 98% of the adult canine muscle cells were positive for CD56 by flow cytometry. These cells expressed MyoD and myogenin as determined by RT-PCR, and desmin as determined by immunocytochemistry. Our method proved to be convenient and practical with a high probability of success. Studies of cell therapy using highly purified myoblasts may now be broadly applied to canine models of human muscle and non-muscle diseases.

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