Abstract
SummaryThe generation of inducible pluripotent stem cells (iPSCs) is a revolutionary technique allowing production of pluripotent patient-specific cell lines used for disease modeling, drug screening, and cell therapy. Integrity of nuclear DNA (nDNA) is mandatory to allow iPSCs utilization, while quality control of mitochondrial DNA (mtDNA) is rarely included in the iPSCs validation process. In this study, we performed mtDNA deep sequencing during the transition from parental fibroblasts to reprogrammed iPSC and to differentiated neuronal precursor cells (NPCs) obtained from controls and patients affected by mitochondrial disorders. At each step, mtDNA variants, including those potentially pathogenic, fluctuate between emerging and disappearing, and some having functional implications. We strongly recommend including mtDNA analysis as an unavoidable assay to obtain fully certified usable iPSCs and NPCs.
Highlights
The exponential increase of applications of induced pluripotent stem cells includes generation of differentiated cells, development of organoids for investigations of disease mechanisms and drug discovery (Shi et al, 2017), and their clinical use for therapeutic purposes in humans (Barker et al, 2017)
We present our own analysis of induced pluripotent stem cells (iPSCs) generated from fibroblasts and peripheral blood mononuclear cells (PBMCs) and, for the first time, of neuronal precursor cells (NPCs)
Variants were analyzed with PhyloTree to reconstruct the haplogroup of each cell line and the full consistency of haplotypes between parental fibroblasts, iPSCs derived clones, and correlated NPCs (Table 1)
Summary
The exponential increase of applications of induced pluripotent stem cells (iPSCs) includes generation of differentiated cells, development of organoids for investigations of disease mechanisms and drug discovery (Shi et al, 2017), and their clinical use for therapeutic purposes in humans (Barker et al, 2017) This poses specific questions on their quality control, and there are concerns about age-related burden of somatic mutations (Kang et al, 2016; Lo Sardo et al, 2017), lineage-specific epigenetic memory affecting the methylation pattern (Nashun et al, 2015), and pre-existing or reprogramming-related occurrence of tumorigenic mutations (Ben-David and Benvenisty, 2011; Gore et al, 2011). Of particular relevance is the potential functional reflection on iPSCs of mtDNA sequence diversity characterizing human populations (Gomez-Duran et al, 2010; Strobbe et al, 2018)
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