We established quail clonal heart muscle cell lines from cardiac rhabdomyosarcomas developed in embryos injected in ovo with the MC29 virus containing the v-myc oncogene. These clones were characterized by means of antibodies detecting markers of striated muscle cells. Two clones were selected for further characterization on the basis of a distribution of myogenic markers similar to that in normal early embryonic cardiac muscle cells. However, these muscle markers progressively disappeared with time in culture. Cardiomyocytic differentiation could be reinduced in culture, by associating the avain cardiac cells with 3T3 cells in a defined synthetic medium. Muscle markers were then reexpressed in all cardiac cells as soon as Day 1 after coculture. Multiplication of cardiac cells continued at the same time. This is characteristic of cardiac clones since MC29-infected quail myoblasts and MC29-infected quail fibroblasts exhibited a split response to 3T3 association, i.e., decreased growth and enhanced differentiation. The cardiac clones were maintained in vitro for more than 60 generations (6 months) without morphological changes. To our knowledge, this is the first description of clonal embryonic avian heart cell lines.