Counting stem cells: methodological constraints

Leonid V. Bystrykh, Evgenia Verovskaya, Erik Zwart, Mathilde Broekhuis, Gerald de Haan*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

66 Citations (Scopus)

Abstract

The number of stem cells contributing to hematopoiesis has been a matter of debate. Many studies use retroviral tagging of stem cells to measure clonal contribution. Here we argue that methodological factors can impact such clonal analyses. Whereas early studies had low resolution, leading to underestimation, recent methods may result in an overestimation of stem-cell counts. We discuss how restriction enzyme choice, PCR bias, high-throughput sequencing depth and tagging method could affect the conclusions of clonal studies.

Original languageEnglish
Pages (from-to)567-574
Number of pages8
JournalNature Methods
Volume9
Issue number6
DOIs
Publication statusPublished - Jun-2012

Keywords

  • SCID-X1 GENE-THERAPY
  • ADENOSINE-DEAMINASE DEFICIENCY
  • INTEGRATION SITE ANALYSIS
  • VECTOR INTEGRATION
  • RETROVIRAL INTEGRATION
  • HEMATOPOIETIC SYSTEM
  • PERIPHERAL-BLOOD
  • PROGENITOR CELLS
  • CD34(+) CELLS
  • CORD BLOOD

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