title: The Number of Point Mutations in Induced Pluripotent Stem Cells and Nuclear Transfer Embryonic Stem Cells Depends on the Method and Somatic Cell Type Used for Their Generation.
abstract: Induced pluripotent stem cells hold great promise for regenerative medicine but point mutations have been identified in these cells and have raised serious concerns about their safe use. We generated nuclear transfer embryonic stem cells (ntESCs) from both mouse embryonic fibroblasts (MEFs) and tail-tip fibroblasts (TTFs) and by whole genome sequencing found fewer mutations compared with iPSCs generated by retroviral gene transduction. Furthermore, TTF-derived ntESCs showed only a very small number of point mutations, approximately 80% less than the number observed in iPSCs generated using retrovirus. Base substitution profile analysis confirmed this greatly reduced number of point mutations. The point mutations in iPSCs are therefore not a Yamanaka factor-specific phenomenon but are intrinsic to genome reprogramming. Moreover, the dramatic reduction in point mutations in ntESCs suggests that most are not essential for genome reprogramming. Our results suggest that it is feasible to reduce the point mutation frequency in iPSCs by optimizing various genome reprogramming conditions. We conducted whole genome sequencing of ntES cells derived from MEFs or TTFs. We thereby succeeded in establishing TTF-derived ntES cell lines with far fewer point mutations. Base substitution profile analysis of these clones also indicated a reduced point mutation frequency, moving from a transversion-predominance to a transition-predominance. Stem Cells 2017;35:1189-1196.
authors.affiliation: Department of Basic Medical Sciences for Radiation Damages, National Institute of Radiological Sciences, National Institutes for Quantum and Radiological Science and Technology, Chiba, Japan.
authors.forename: Ryoko
authors.initials: R
authors.lastname: Araki
authors.affiliation: Department of Biotechnology, Faculty of Life and Environmental Science, University of Yamanashi, Kofu, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Mizutani
authors.affiliation: Department of Basic Medical Sciences for Radiation Damages, National Institute of Radiological Sciences, National Institutes for Quantum and Radiological Science and Technology, Chiba, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Hoki
authors.affiliation: Department of Basic Medical Sciences for Radiation Damages, National Institute of Radiological Sciences, National Institutes for Quantum and Radiological Science and Technology, Chiba, Japan.
authors.forename: Misato
authors.initials: M
authors.lastname: Sunayama
authors.affiliation: Department of Biotechnology, Faculty of Life and Environmental Science, University of Yamanashi, Kofu, Japan.
authors.forename: Sayaka
authors.initials: S
authors.lastname: Wakayama
authors.affiliation: Department of Biotechnology, Faculty of Life and Environmental Science, University of Yamanashi, Kofu, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nagatomo
authors.affiliation: Department of Basic Medical Sciences for Radiation Damages, National Institute of Radiological Sciences, National Institutes for Quantum and Radiological Science and Technology, Chiba, Japan.
authors.forename: Yasuji
authors.initials: Y
authors.lastname: Kasama
authors.affiliation: Department of Basic Medical Sciences for Radiation Damages, National Institute of Radiological Sciences, National Institutes for Quantum and Radiological Science and Technology, Chiba, Japan.
authors.forename: Miki
authors.initials: M
authors.lastname: Nakamura
authors.affiliation: Department of Biotechnology, Faculty of Life and Environmental Science, University of Yamanashi, Kofu, Japan.
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Wakayama
authors.affiliation: Department of Basic Medical Sciences for Radiation Damages, National Institute of Radiological Sciences, National Institutes for Quantum and Radiological Science and Technology, Chiba, Japan.
authors.forename: Masumi
authors.identifier: 0000-0003-1550-2006
authors.initials: M
authors.lastname: Abe
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D004622:Embryo, Mammalian / Q000166:cytology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005787:Gene Frequency / Q000235:genetics; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008810:Mice, Inbred C57BL; D053652:Nuclear Transfer Techniques; D016366:Open Reading Frames / Q000235:genetics; D017354:Point Mutation / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D017422:Sequence Analysis, DNA; D013623:Tail
keywords: Genetic aberrations; Genome reprogramming; Induced pluripotent stem cells; Nuclear transfer embryonic stem cell; Unique base substitution profile; Whole genome sequencing