SV40 immortalization of adult human mesenchymal cells from neuroretina. Biological, functional and molecular characterization.

Human adult mesenchymal cells from neuroretina (human choroid cells, HC) have acquired an infinite lifespan, following phenotypic transformation with a wild-type SV40. Immortalized cells (HC/SV40) contain high numbers of free circular viral DNA, and integrated molecules in a head-to-tail array in the cellular DNA. HC/SV40 cells express both the virus-coded "T" antigens and the cell-coded p53 transformation-associated protein. The transformed phenotype was further characterized by loss of contact inhibition of cell division, inability to induce the retraction of a fibrin clot and to spread within fibrin, and the existence of an altered distribution of actin cables. For the first time we also describe a coupling of the immunofluorescence and the quantitative cytofluorometric analyses, a new transformation parameter, since we show that SV40 transformation causes reorganization of the cell membrane by inducing the unmasking of the antigen recognized by the 4F2 monoclonal antibody, which is present in a "cryptic" form in the untransformed cells. Though the HC/SV40 cells have been continuously passaged over a 3-year period, they have not yet achieved a fully malignant phenotype, since they retain serum-dependency and the presence of a well developed fibronectin pericellular network, and they are not tumorigenic in nude mice. Thus this human immortal cell line constitutes a very useful tool for studying the progression toward full malignancy and the relationships between evolution of transformation parameters and changes in the viral and cellular genome interplay.