Yeoh J S G, Ausema A, Wierenga P, de Haan G, van Os R
Department of Cell Biology, Section Stem Cell Biology, University Medical Centre Groningen, Groningen, The Netherlands.
Bone Marrow Transplant. 2007 Apr;39(7):401-9. doi: 10.1038/sj.bmt.1705601. Epub 2007 Feb 12.
In this study, we use competitive repopulation to compare the quality and frequency of stem cells isolated from mobilized blood with stem cells isolated from bone marrow (BM) in a mouse model. Lin(-)Sca-1(+)c-Kit(+) (LSK) cells were harvested from control BM and peripheral blood of mice following granulocyte colony-stimulating factor (G-CSF) administration. LSK cells were used because of their resemblance to human CD34(+) cells. We confirmed that transplantation of phenotypically defined mobilized peripheral blood (MPB) stem cells results in rapid recovery of blood counts. However, in vitro results indicated that LSK cells purified from MPB had lower cobblestone area-forming cell day 35 activity compared to BM. Additionally, evaluation of chimerism after co-transplantation of LSK cells purified from blood and BM revealed that MPB stem cells contained 25-fold less repopulation potential compared to BM stem cells. Competitive repopulating unit frequency analysis showed that freshly isolated MPB LSK cells have 8.8-fold fewer cells with long-term repopulating ability compared to BM LSK cells. Secondary transplantation showed no further decline in contribution of hematopoiesis relative to BM. We conclude that the reduced frequency of stem cells within the LSK population of MPB, rather than poorer quality, causes the reduced repopulation potential.Bone Marrow Transplantation (2007) 39, 401-409. doi:10.1038/sj.bmt.1705601; published online 12 February 2007.
在本研究中,我们利用竞争性再增殖,在小鼠模型中比较从动员血液中分离的干细胞与从骨髓(BM)中分离的干细胞的质量和频率。在给予粒细胞集落刺激因子(G-CSF)后,从小鼠的对照骨髓和外周血中收获Lin(-)Sca-1(+)c-Kit(+)(LSK)细胞。使用LSK细胞是因为它们与人类CD34(+)细胞相似。我们证实,表型明确的动员外周血(MPB)干细胞移植可使血细胞计数迅速恢复。然而,体外结果表明,与骨髓相比,从MPB中纯化的LSK细胞在第35天形成鹅卵石区域的细胞活性较低。此外,对从血液和骨髓中纯化的LSK细胞进行共移植后的嵌合情况评估显示,与BM干细胞相比,MPB干细胞的再增殖潜力低25倍。竞争性再增殖单位频率分析表明,与BM LSK细胞相比,新鲜分离的MPB LSK细胞中具有长期再增殖能力的细胞数量少8.8倍。二次移植显示,相对于骨髓,造血贡献没有进一步下降。我们得出结论,MPB的LSK群体中干细胞频率降低而非质量较差导致了再增殖潜力降低。《骨髓移植》(2007年)39卷,401 - 409页。doi:10.1038/sj.bmt.1705601;2007年2月12日在线发表。