Detecting Genetic Mosaicism in Cultures of Human Pluripotent Stem Cells

Stem Cell Reports. 2016 Nov 8;7(5):998-1012. doi: 10.1016/j.stemcr.2016.10.003.


Genetic changes in human pluripotent stem cells (hPSCs) gained during culture can confound experimental results and potentially jeopardize the outcome of clinical therapies. Particularly common changes in hPSCs are trisomies of chromosomes 1, 12, 17, and 20. Thus, hPSCs should be regularly screened for such aberrations. Although a number of methods are used to assess hPSC genotypes, there has been no systematic evaluation of the sensitivity of the commonly used techniques in detecting low-level mosaicism in hPSC cultures. We have performed mixing experiments to mimic the naturally occurring mosaicism and have assessed the sensitivity of chromosome banding, qPCR, fluorescence in situ hybridization, and digital droplet PCR in detecting variants. Our analysis highlights the limits of mosaicism detection by the commonly employed methods, a pivotal requirement for interpreting the genetic status of hPSCs and for setting standards for safe applications of hPSCs in regenerative medicine.

Keywords: Human pluripotent stem cells; detection methods; genetic changes; sensitivity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques
  • Cell Line
  • Chromosomes, Human
  • Chromosomes, Human, Pair 17
  • Chromosomes, Human, Pair 20
  • DNA Copy Number Variations
  • Genetic Variation*
  • Humans
  • In Situ Hybridization, Fluorescence
  • Karyotype
  • Mosaicism*
  • Pluripotent Stem Cells / cytology
  • Pluripotent Stem Cells / metabolism*
  • Polymerase Chain Reaction
  • Trisomy

Supplementary concepts

  • Chromosome 17 trisomy