Quantitative mitochondrial DNA copy number determination using droplet digital PCR with single cell resolution: a focus on aging and cancer

https://www.biorxiv.org/content/biorxiv/early/2019/03/16/579789.full.pdf

Ryan O’Hara, Enzo Tedone,, Andrew Ludlow, Ejun Huang, Beatrice Arosio, Daniela Mari, Jerry W. Shay

• The authors develop a protocol to measure single-cell mtDNA copy number using digital droplet PCR
• The authors find ~10-fold inter-cellular variability in mtDNA copy number (in an immortalised human cell line, H1299), which could not be fully explained by cell cycle variations.
• The authors investigated how mtDNA copy number changes after stimulation of peripheral blood mononuclear cells (PBMCs, a heterogeneous cellular population largely consisting of T cells). Previous studies have shown a decline in mtDNA copy number with ageing in this cell population. The authors studied stimulated PBMCs in young, old, and healthy/frail centenarians. Healthy centenarians tended to have higher mtDNA copy number than frail, or ~70 year old, individuals.