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 |
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Pages (from-to) | 567-574 |
Number of pages | 8 |
Journal | Nature Methods |
Volume | 9 |
Issue number | 6 |
DOIs | |
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