Detecting Genetic Mosaicism in Cultures of Human Pluripotent Stem Cells

Stem Cell Reports

Thomas F. Allison¹  Adam J. Hirst¹  Paul J. Gokhale¹  Harry D. Moore¹  Peter W. Andrews¹  Ivana Barbaric¹  Duncan Baker¹  Miguel A. Juarez²  Mark Wheeler³  Kerry Bean³  Steve Williams³ 

[1] Department of Biomedical Science, Centre for Stem Cell Biology, The University of Sheffield, Sheffield S10 2TN, UK;School of Mathematics and Statistics, The University of Sheffield, Sheffield S3 7RH, UK;Sheffield Diagnostic Genetic Services, Sheffield Children’s Hospital, Sheffield S10 2TH, UK;

关键词: Human pluripotent stem cells;    genetic changes;    detection methods;    sensitivity;   

DOI  :  10.1016/j.stemcr.2016.10.003

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown