Studies on the effects of time and passage on porcine primary muscle cell cultures and methods to purify myoblasts were conducted using flow cytometry and fluorescence-activated cell sorting (FACS). Primary muscle cells cultured on single plates revealed a small cell (<10 mm diameter) population consisting of 90% desmin-positive myoblasts and a large cell (> or = 10 mm diameter) population containing desmin-positive myoblasts and nonmyoblasts. The small myoblasts were detectable up to 28 days but after cell sorting and passage, they became indistinguishable from the large myoblast population. This indicates that pig muscle contains small self-renewing myoblasts similar to humans, that become larger when induced to proliferate. A human myoblast-specific monoclonal antibody allows FACS of both large and small myoblasts from primary cells within 2 days of culture and independent of passage. These characteristics of porcine myoblasts indicate that the pig may be a suitable large animal model for myoblast-mediated gene transfer.