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Detecting Genetic Mosaicism in Cultures of Human Pluripotent Stem Cells.

Abstract：

: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.

journal_name

Stem Cell Reports

journal_title

Stem cell reports

authors

Baker D,Hirst AJ,Gokhale PJ,Juarez MA,Williams S,Wheeler M,Bean K,Allison TF,Moore HD,Andrews PW,Barbaric I

doi

10.1016/j.stemcr.2016.10.003

subject

Has Abstract

pub_date

2016-11-08 00:00:00

pages

998-1012

issue

5

issn

2213-6711

pii

S2213-6711(16)30241-7

journal_volume

7

pub_type

杂志文章

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