Whole-genome sequencing and mutational analysis of human cord-blood derived stem and progenitor cells

Axel Rosendahl Huber, Anaïs J.C.N. van Leeuwen, Flavia Peci, Jurrian K. de Kanter, Eline J.M. Bertrums, Ruben van Boxtel*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

1 Citation (Scopus)
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Abstract

Mutational signatures have been identified in cancer genomes, providing information about the causes of cancer and treatment vulnerabilities. This protocol describes an assay to determine the genotoxic mechanisms underlying these signatures using cord-blood derived hematopoietic stem and progenitor cells (CB-HSPCs). CB-HSPCs have a low mutation background, enabling sensitive detection of mutations. First, CB-HSPCs are exposed in vitro, sorted, and clonally expanded. This expansion enables whole-genome sequencing to detect the mutation load and respective patterns induced during genotoxic exposure. For complete details on the use and execution of this protocol, please refer to de Kanter et al. (2021).

Original languageEnglish
Article number101361
JournalSTAR Protocols
Volume3
Issue number2
Early online date6 May 2022
DOIs
Publication statusPublished - 17 Jun 2022

Bibliographical note

Funding Information:
This research was supported by funding from a NWO Vidi grant to R.v.B, no. 016.Vidi.171.023, and the Oncode Institute . The authors want to thank M.A. Satzl for providing pictures of the CMBC isolation.

Funding Information:
This research was supported by funding from a NWO Vidi grant to R.v.B, no. 016.Vidi.171.023, and the Oncode Institute. The authors want to thank M.A. Satzl for providing pictures of the CMBC isolation. A.R.H. and A.J.C.N.v.L. performed the experiments. A.R.H. performed the bioinformatic analyses. A.J.C.N.v.L, A.R.H. F.P, E.J.M.B, and J.K.d.K. wrote the manuscript. All authors read, revised, and approved the manuscript. A.R.H. A.v.L. and R.v.B. are named as inventors on a patent application filed resulting from this work.

Publisher Copyright:
© 2022 The Author(s)

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