An established avian fibroblast cell line without mitochondrial DNA.
An established avian fibroblast cell line (LSCC-H32) has been found to be inherently resistant to the growth-inhibitory effect of ethidium bromide, when supplied with exogenous uridine. After long-term exposure to ethidium bromide (90 days), the cell population has been transferred to drug-free medium for 60 days, and then seeded at low cell density. Three clones have been isolated and propagated in drug-free medium for 5, 6, and more than 12 months, respectively. It was found that none of these cell lines had detectable cytochrome c oxidase activity and that they were virtually devoid of cytochromes aa3 and b. Mitochondrial DNA was quantitated by DNA-DNA reassociation kinetics with a probe of chicken liver mitochondrial DNA. A mean number of 300 copies of mitochondrial DNA per cell was found in LSCC-H32 cells. Analysis of DNA extracted from cell populations exposed to ethidium bromide for 90 days and then transferred to drug-free medium for long periods of time revealed no mitochondrial DNA molecules by reassociation kinetics or by Southern blot hybridization of HindIII-or AvaI-digested total cellular DNA.
D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D046508:Culture Techniques / Q000379:methods; D003580:Cytochromes / Q000032:analysis; D004272:DNA, Mitochondrial / Q000032:analysis* / Q000235:genetics; D003576:Electron Transport Complex IV / Q000032:analysis; D004996:Ethidium / Q000494:pharmacology; D005347:Fibroblasts / Q000032:analysis; D008099:Liver / Q000032:analysis; D009693:Nucleic Acid Hybridization; D011506:Proteins / Q000032:analysis
D003580:Cytochromes; D004272:DNA, Mitochondrial; D011506:Proteins; D003576:Electron Transport Complex IV; D004996:Ethidium