CELL LINES
Contributor Information
- Name Payam Gammage
- Institute Cancer Research UK, Glasgow: The Beatson Institute
Tool Details
- Tool name: 143B m.8993T>G isogenic mTUNE mT80 Cell Line
- Alternate names: mTUNE 8, mT8
- Tool type: Cell Lines
- Tool sub-type: Continuous
- Parental cell line: 143B human osteosarcoma (m.8993T>G cybrid)
- Organism: Human
- Tissue: Bone
- Cancer type: Sarcoma
- Disease: Cancer
- Model: Tumour line
- Description: The cellular metabolic environemnt is influenced by relative levels of coexistant mutant and wild-type mitochondrial DNAs (heteorplasmy) in a range of diseases for which mitochondrial dysfunction is a feature. Disease phenotypes may be rescued by returning heteroplasmy towards wild-type levels. To investigate this approach, the team led by Payam Gammage have devised a series of cell lines which stably express stable levels of mutatnt m.8993T>G mtDNA heteroplasmy. Cell lines bearing m.8993T>G heteroplasmy of 80, 45, and 10% are available at CancerTools.org. These lines are derivatives of the m.8993T cybrid cell developed by Prof Eric Schon, and are of of 143B human osteosarcoma lineage. This is part of a series of three cell line (mTUNE); see Related research tools tab.
- Research area: Cancer; Metabolism
- Production details: mTUNE cells were generated by Dr Michal MinczukÄË?Â?Âs lab and derive from female human osteosarcoma 143B (RRID: CVCL_2270) cybrid cells (Porteous et al., 1998), after correction of m.8993T>G mutation with mitochondrially-targeted zinc finger nucleases (Gammage et al., 2016a). See Gaude et al., 2018 for further details.
- Additional notes: This is part of a series of three cell line (mTUNE); see Related Reagents tab.
- For Research Use Only
Target Details
Application Details
Handling
- Format: Frozen
- Growth medium: high glucose DMEM media supplemented with L-glutamine and 10%FBS and pen/strep
- Shipping conditions: Dry ice
- Subculture routine: Grow in regular dishes or flasks (10cm dish or 75cm2 flask). Passage 2 or 3 times a week, split when confluent (wash with PBS and then use trypsin to split). There is no maximum passage number. 1:10 for 3 splits a week, 1:20 Friday to Monday
Documentation
References
- • Genome editing in mitochondria corrects a pathogenic mtDNA mutation in vivo.
- • Enhanced Manipulation of Human Mitochondrial DNA Heteroplasmy In Vitro Using Tunable mtZFN Technology.
- • NADH Shuttling Couples Cytosolic Reductive Carboxylation of Glutamine with Glycolysis in Cells with Mitochondrial Dysfunction.
- • Engineered mtZFNs for Manipulation of Human Mitochondrial DNA Heteroplasmy.
- • Near-complete elimination of mutant mtDNA by iterative or dynamic dose-controlled treatment with mtZFNs.
- • Mitochondrially targeted ZFNs for selective degradation of pathogenic mitochondrial genomes bearing large-scale deletions or point mutations.
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