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Concise Review: Assessing the Genome Integrity of Human Induced Pluripotent Stem Cells: What Quality Control Metrics?

Assou, Said, Bouckenheimer, Julien, De Vos, John·2018·Stem Cells
DOI 10.1002/stem.2797
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Abstract

Human induced pluripotent stem cells (hiPSCs) have the potential to differentiate virtually into any cell type in unlimited quantities. Therefore, they are ideal for in vitro tissue modeling or to produce cells for clinical use. Importantly, and differently from immortalized and cancer cell lines, the hiPSC genome scrupulously reproduces that of the cell from which they were derived. However, hiPSCs can develop genetic abnormalities during reprogramming or prolonged cell culture, such as aneuploidies or oncogenic mutations (e.g., in TP53). Therefore, hiPSC genome integrity must be routinely monitored because serious genome alterations would greatly compromise their usefulness or safety of use. Here, we reviewed hiPSC genome quality control monitoring methods and laboratory practice. Indeed, due to their frequency and functional consequences, recurrent genetic defects found in cultured hiPSCs are inacceptable and their appearance should be monitored by routine screening. Hence, for research purposes, we propose that the genome of hiPSC lines should be systematically screened at derivation, at least by karyotyping, and then regularly (every 12 weeks) during experiments, for instance with polymerase chain reaction-based techniques. For some specific applications, such as research on aging, cell cycle, apoptosis or cancer, other tests (e.g., TP53 mutation detection) should also be included. For clinical use, in addition to karyotyping, we advise exome sequencing.

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  • Such genetic changes appear to occur at any time during passage and may be associated with culture conditions such as medium acidification, frequent single cell dissociation, or repeated use of Rho kinase inhibitors.

    “Such changes appear to occur at any time during passage (International Stem Cell Initiative, 2011) and may be associated with culture conditions - such as medium acidification - as a consequence of suboptimal culturing and passaging (e.g., frequent single cell dissociation) or the repeated use of Rho kinase inhibitors (Assou et al., 2018; Jacobs et al., 2016).”

    From Guidance document on Good cell and tissue culture practice 2.0 (GCCP 2.0) (2021) · cites this paper as Assou et al., 2018

    In ‘Classification According to the Cause’ (page not provided), the review links suboptimal culture and single-cell passaging to genomic abnormalities. Our copy does not link medium acidification or repeated use of Rho kinase inhibitors to those changes; its mention of ROCK concerns apoptosis during single-cell dissociation. The ‘any time during passage’ clause is attributed to a different citation in the printed sentence. (LLM, conf=0.86)

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Cites

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Cited by

Sorted by citations

Authors
Assou, S., Bouckenheimer, J., & De Vos, J.
Affiliations
IRMB, Univ Montpellier, INSERM, CHU Montpellier, Montpellier, France
ORCID
De Vos, John
Venue
Stem Cells
Publisher
Oxford University Press (OUP)
Published
2018-02-18
Volume
36
Issue
6
Pages
814-821
ISSN
  • 1066-5099
  • 1549-4918
DOI
10.1002/stem.2797
Cited by
74
References
13
Language
en
Licence
academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)

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