Few single nucleotide variations in exomes of human cord blood induced pluripotent stem cells

PLoS One. 2013;8(4):e59908. doi: 10.1371/journal.pone.0059908. Epub 2013 Apr 1.

Abstract

The effect of the cellular reprogramming process per se on mutation load remains unclear. To address this issue, we performed whole exome sequencing analysis of induced pluripotent stem cells (iPSCs) reprogrammed from human cord blood (CB) CD34(+) cells. Cells from a single donor and improved lentiviral vectors for high-efficiency (2-14%) reprogramming were used to examine the effects of three different combinations of reprogramming factors: OCT4 and SOX2 (OS), OS and ZSCAN4 (OSZ), OS and MYC and KLF4 (OSMK). Five clones from each group were subject to whole exome sequencing analysis. We identified 14, 11, and 9 single nucleotide variations (SNVs), in exomes, including untranslated regions (UTR), in the five clones of OSMK, OS, and OSZ iPSC lines. Only 8, 7, and 4 of these, respectively, were protein-coding mutations. An average of 1.3 coding mutations per CB iPSC line is remarkably lower than previous studies using fibroblasts and low-efficiency reprogramming approaches. These data demonstrate that point nucleotide mutations during cord blood reprogramming are negligible and that the inclusion of genome stabilizers like ZSCAN4 during reprogramming may further decrease reprogramming-associated mutations. Our findings provide evidence that CB is a superior source of cells for iPSC banking.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Dedifferentiation
  • Cells, Cultured
  • Coculture Techniques
  • DNA Mutational Analysis
  • Exome
  • Fetal Blood / cytology*
  • Genomic Instability
  • Humans
  • Induced Pluripotent Stem Cells / physiology*
  • Induced Pluripotent Stem Cells / transplantation
  • Kruppel-Like Factor 4
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Open Reading Frames
  • Point Mutation*
  • Regenerative Medicine
  • Teratoma / pathology

Grants and funding

This research was supported by Loma Linda University Department of Medicine (XBZ), and DOD Concept Award W81XWH-11-1-0607 (XBZ), the Department of Pathology and Human Anatomy and the Center for Health Disparities and Molecular Medicine at Loma Linda University (KJP and RJS). This work was also supported by the “863 Projects” of Ministry of Science and Technology of China (2011AA020118 to XF) and the “Strategic Priority Research Program” of the Chinese Academy of Sciences, Stem Cell and Regenerative Medicine Research (XDA01040405 to XF). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.