Systematic determination of the mitochondrial proportion in human and mice tissues for single-cell RNA sequencing data quality control Academic Article uri icon


  • Motivation Quality control (QC) is a critical step in single-cell RNA-seq (scRNA-seq) data analysis. Low-quality cells are removed from the analysis during the QC process to avoid misinterpretation of the data. One of the important QC metrics is the mitochondrial proportion (mtDNA%), which is used as a threshold to filter out low-quality cells. Early publications in the field established a threshold of 5% and since then, it has been used as a default in several software packages for scRNA-seq data analysis and adopted as a standard in many scRNA-seq studies. However, the validity of using a uniform threshold across different species, single-cell technologies, tissues, and cell types has not been adequately assessed.Results We systematically analyzed 5,530,106 cells reported in 1,349 annotated datasets available in the PanglaoDB database and found that the average mtDNA% in scRNA-seq data across human tissues is significantly higher than in mouse tissues. This difference is not confounded by the platform used to generate the data. Based on this finding, we propose new reference values of the mtDNA% for 121 tissues of mice and 44 tissues of humans. In general, for mouse tissues, the 5% threshold performs well to distinguish between healthy and low-quality cells. However, for human tissues, the 5% threshold should be reconsidered as it fails to accurately discriminate between healthy and low-quality cells in 29.5% (13 of 44) tissues analyzed. We conclude that omitting the mtDNA% QC filter or adopting a suboptimal mtDNA% threshold may lead to erroneous biological interpretations of scRNA-seq data.Availability The code used to download datasets, perform the analyzes, and produce the figures is available at dcosorioh{at}tamu.eduSupplementary information Supplementary data are available at Bioinformatics online.Competing Interest StatementThe authors have declared no competing interest.

author list (cited authors)

  • Osorio, D., & Cai, J. J.

complete list of authors

  • Osorio, D||Cai, JJ

publication date

  • January 1, 2020 11:11 AM