Isolation of two populations of myoblasts from porcine skeletal muscle

Studies on the effects of time and passage on porcine primary muscle cell c ultures and methods to purify myoblasts were conducted using flow cytometry and fluorescence-activated cell sorting (FACS). Primary muscle cells cultu red on single plates revealed a small cell (<10 mm diameter) population con sisting of 90% desmin-positive myoblasts and a large cell (greater than or equal to 10 mm diameter) population containing desmin-positive myoblasts an d nonmyoblasts. The small myoblasts were detectable up to 28 days but after cell sorting and passage, they became indistinguishable from the large myo blast population. This indicates that pig muscle contains small self-renewi ng myoblasts similar to humans, that become larger when induced to prolifer ate. A human myoblast-specific monoclonal antibody allows FAGS of both larg e and small myoblasts from primary cells within 2 days of culture and indep endent of passage. These characteristics of porcine myoblasts indicate that the pig may be a suitable large animal model for myoblast-mediated gene tr ansfer. (C) 1999 John Wiley & Sons, Inc.