Sensitivities of NIH/3T3-derived Clonal Cell Lines to Ionizing Radiation: Significance for Gene Transfer Studies1

Rodent cells are frequently used as recipients in experiments involving gene transfer, isolation, and characterization. The present studies were designed to investigate the clonal responses to ionizing radiation of Mil/ 3T3 cells subjected to DNA-mediated gene transfer. Radiation sensitivity (Do) values were determined for the parental NIH/3T3 cell strain, six clonal cell lines transfected with DNA from radiation-resistant human tumor cells, and six nontransfected clonal cell lines. The radiation sen sitivities of four transfected and two nontransfected clonal cell lines differed significantly from parental NIH/3T3 cells (/' < 0.05). Detailed karyotype analysis of two nontransfected clonal cell lines with differing radiation sensitivities showed variation in chromosomal composition. Specifically, a minute chromosome was observed to segregate consistently (in 49 of SO metaphases) with the genome of one NIH/3T3 clone (Mi 2.07 Gy) and was completely absent (from 50 metaphases) in another NIH/3T3 clone (A> 1.06 Gy). In the parental NIH/3T3 strain (A, 2.02 Gy) 10% of cells (3 of 30 metaphases) had such minute chromosomes. These findings demonstrate that the clonal cellular heterogeneity of Mil/ 3T3 cells is characterized by genotypic and phenotypic variations which must be considered in the experimental design involving gene transfer and expression.