Rhabdomyosarcoma rho(0) cells: isolation and characterization of a mitochondrial DNA depleted cell line with 'muscle-like' properties.

TitleRhabdomyosarcoma rho(0) cells: isolation and characterization of a mitochondrial DNA depleted cell line with 'muscle-like' properties.
Publication TypeJournal Article
Year of Publication2000
AuthorsVergani, L, Prescott, AR, Holt, IJ
JournalNeuromuscul Disord
Date Published2000 Aug
KeywordsAntigens, Differentiation, Cell Differentiation, Cell Division, Creatine Kinase, DNA, Mitochondrial, Ethidium, Humans, Rhabdomyosarcoma, Tetradecanoylphorbol Acetate, Tumor Cells, Cultured

Mutations of mitochondrial DNA are a significant cause of neuromuscular disease. Pathological mutant mitochondrial DNA has been studied in control nuclear backgrounds. These experiments entailed transfer of patient-derived mitochondria to rho(0) cells that lack mtDNA. A limitation of these studies has been the fact that the control nuclear backgrounds were unrelated to the affected tissues of patients. Therefore a rhabdomyosarcoma cell line that has 'muscle-like' properties was tested to determine whether it could be depleted of mtDNA. A human rhabdomyosarcoma cell line was treated with the DNA intercalating dye ethidium bromide (3, 8-diamino-5-ethyl-6-phenylphenanthridinium bromide) for 45 days. The treatment induced complete and permanent loss of mitochondrial DNA (rho(0)) in the rhabdomyosarcoma cells, as mtDNA remained undetectable after 8 months of growth in medium without drug. Crucially, the rhabdomyosarcoma rho(0) cells retained the ability to differentiate into myotubes with expression of muscle specific isoenzymes. The rhabdomyosarcoma rho(0) cell line provides a model system for studying pathological mutant mtDNA in cells that more closely resemble human muscle than the hitherto available human rho(0) cell lines.

Alternate JournalNeuromuscul. Disord.
Citation Key10.1016/s0960-8966(00)00096-1
PubMed ID10899454
Grant List1252 / / Telethon / Italy
MC_U105663140 / / Medical Research Council / United Kingdom