Evaluation of transforming activity of cellular DNAs from different origins by NIH3T3 transfection test.
The NIH3T3 cell transfection test, as first described by Cooper, has been optimized, then used to examine the transforming activity of genomic DNA extracted from eucaryotic cell lines commonly used for preparing vaccines or biopharmaceuticals. Accurate assessment of technical parameters of the test has led to improvement in reproducibility, while the demonstration of dose-effect relationships has allowed the definition of applications and limits for quantitative use. We have performed the direct assessment of transforming activity of cellular DNAs from cell lines widely used in biotechnology. In particular, we have shown that genomic DNA extracted from Vero, CHO or MRC5 cells, as well as from human or murine lymphoid cells, has no detectable transforming activity on NIH3T3 cells. Lastly, it has been demonstrated that acidic pH conditions are sufficient to destroy the major part--if not all--of the transforming activity of positive control DNAs.
Carcagne
J
J
Department of Immunology, Pasteur Mérieux Sérums et Vaccins, Marcy L'Etoile, France.
Ha
K T
KT
Armand
J
J
D016475:3T3 Cells; D000818:Animals; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D004247:DNA / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000302:isolation & purification; D005069:Evaluation Studies as Topic; D005990:Glycerol; D006801:Humans; D006863:Hydrogen-Ion Concentration; D051379:Mice; D009374:Neoplasms, Experimental / Q000209:etiology; D051381:Rats; D011919:Rats, Inbred Strains; D012680:Sensitivity and Specificity; D014162:Transfection; D014170:Transformation, Genetic
D004247:DNA; D005990:Glycerol