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Evidence for a multistep pathogenesis in the generation of tumorigenic cell lines from hemopoietic colonies exposed to Abelson virus in vitro.

The present studies were undertaken to investigate the ability of Abelson murine leukemia virus (A-MuLV) to transform cells derived in vitro from pluripotent hemopoietic progenitor cells of high proliferative potential. We now report that continuously growing, autonomous cell lines could be obtained from a high proportion of individually infected multilineage colonies generated in assays of spleen cells from normal adult mice if the infected cells were cocultivated for the first two to three months with irradiated NIH-3T3 cells. No lines were obtained if the 3T3 cell feeders were not initially present. Similar results were obtained when the cells exposed to virus were from multilineage colonies originating from isolated single cells obtained by replating small blast colonies. Characterization of the transformants and a number of derivative cloned sublines revealed the consistent presence of a mast cell phenotype, with some suggestion of macrophage differentiation in a few cases. All cell lines tested produced virus, showed a variable pattern of A-MuLV integration, and gave rise directly to tumors when injected subcutaneously, as shown by both Southern analysis and cytogenetic studies. The early absolute but transient dependence of these A-MuLV mast cell transformants on a fibroblast feeder suggests a multistep process in their evolution, in which the acquisition of autonomy from factors of mesenchymal cell origin may play an important role.