Laboratory of Stem Cell Research, European Research Institute for the Biology of Ageing, University Medical Center Groningen, University of Groningen, The Netherlands.
Nat Methods. 2012 May 30;9(6):567-74. doi: 10.1038/nmeth.2043.
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.
造血干细胞数量一直存在争议。许多研究使用逆转录病毒标记干细胞来衡量克隆贡献。在这里，我们认为方法学因素会影响此类克隆分析。虽然早期的研究分辨率较低，导致低估，但最近的方法可能会高估干细胞的数量。我们讨论了限制酶选择、PCR 偏倚、高通量测序深度和标记方法如何影响克隆研究的结论。
