Counting stem cells: methodological constraints

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

doi:10.1038/NMETH.2043

Counting stem cells: methodological constraints

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

^*Corresponding author for this work

Faculty of Medical Sciences/UMCG

Research output: Contribution to journal › Article › Academic › peer-review

67 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 language	English
Pages (from-to)	567-574
Number of pages	8
Journal	Nature Methods
Volume	9
Issue number	6
DOIs	https://doi.org/10.1038/NMETH.2043
Publication status	Published - 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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title = "Counting stem cells: methodological constraints",

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.",

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author = "Bystrykh, {Leonid V.} and Evgenia Verovskaya and Erik Zwart and Mathilde Broekhuis and {de Haan}, Gerald",

year = "2012",

month = jun,

doi = "10.1038/NMETH.2043",

language = "English",

volume = "9",

pages = "567--574",

journal = "Nature Methods",

issn = "1548-7091",

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T1 - Counting stem cells

T2 - methodological constraints

AU - Bystrykh, Leonid V.

AU - Verovskaya, Evgenia

AU - Zwart, Erik

AU - Broekhuis, Mathilde

AU - de Haan, Gerald

PY - 2012/6

Y1 - 2012/6

N2 - 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.

AB - 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.

KW - SCID-X1 GENE-THERAPY

KW - ADENOSINE-DEAMINASE DEFICIENCY

KW - INTEGRATION SITE ANALYSIS

KW - VECTOR INTEGRATION

KW - RETROVIRAL INTEGRATION

KW - HEMATOPOIETIC SYSTEM

KW - PERIPHERAL-BLOOD

KW - PROGENITOR CELLS

KW - CD34(+) CELLS

KW - CORD BLOOD

U2 - 10.1038/NMETH.2043

DO - 10.1038/NMETH.2043

M3 - Article

SN - 1548-7091

VL - 9

SP - 567

EP - 574

JO - Nature Methods

JF - Nature Methods

IS - 6

ER -

Counting stem cells: methodological constraints

Abstract

Keywords

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