Growth autonomy and high levels of invasiveness are characteristics of human melanoma cells that are metastatic in vivo. By consecutive passage through a reconstructed basement membrane, we have selected from 5 of 6 primary melanoma cell lines variants which show an up to 10-fold increase in invasiveness. The invasive variants grew more rapidly than the parental, noninvasive cells in serum- and growth factor-free medium and one of the 3 variant cell lines with the highest invasive capacity in vitro metastasized to the lungs when injected s.c. into nude mice. In a second approach, variants of 6 primary melanoma cell lines were clonally selected in medium without exogenous growth factors (protein-free medium). These selected cells showed higher invasive properties in vitro and in vivo than the parental cells. Clones of invasive and growth factor-independent cell variants were heterogenous and changed over time in the absence of selected pressure to a phenotype similar to that of parental nonselected cells. These results indicate that primary melanoma cells contain subpopulations of cells that have the phenotype of an advanced (metastatic) stage of tumor progression, but this phenotype is not stable without selective pressure.