Hematopoietic stem cells do not engraft with absolute efficiencies
Hematopoietic stem cells do not engraft with absolute efficiencies
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DOI：
10.1182/blood-2005-02-0655
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发表时间：
2006-01-15
期刊：
影响因子：
20.3
通讯作者：
Goodell, MA
中科院分区：
文献类型：
--
作者：
Camargo, FD;Chambers, SM;Goodell, MA
造血干细胞（HSC）可以通过其流出Hoechst 33342染料的能力从鼠骨髓中分离。这种方法定义了一个非常小的和造血功能强大的细胞亚群，称为侧群（SP）。最近的研究表明，移植的单个SP细胞能够以接近绝对效率的淋巴造血重建。在这里，我们仔细重新评估单个SP细胞的造血潜力，发现重建率大大降低。我们的策略是将单个So细胞沿着不同群体的竞争细胞（其自我更新能力各不相同）共同移植。即使在最小化HSC竞争的情况下，SP细胞也只能重建高达35%的受体小鼠。此外，通过免疫表型分析和体外克隆试验，我们发现SP细胞几乎是同质的。基于Hoechst排斥和单个细胞表面标记物的HSC分离允许与用复杂的免疫策略获得的富集水平相似的富集水平。总之，我们的研究结果表明，即使是一个非常同质的HSC人口，基于表型和染料流出，不能重建小鼠在绝对的效率。
Hematopoietic stem cells (HSCs) can be isolated from murine bone marrow by their ability to efflux the Hoechst 33342 dye. This method defines an extremely small and hematopoietically potent subset of cells known as the side population (SP). Recent studies suggest that transplanted single SP cells are capable of lymphohematopoietic repopulation at near absolute efficiencies. Here, we carefully reevaluate the hematopoletic potential of individual SP cells and find substantially lower rates of reconstitution. Our strategy involved the cotransplantation of single So cells along with different populations of competitor cells that varied in their self-renewal capacity. Even with minimized HSC competition, SP cells were only able to reconstitute up to 35% of recipient mice. Furthermore, through immunophenotyping and clonal in vitro assays we find that SP cells are virtually homogeneous. Isolation of HSCs on the basis of Hoechst exclusion and a single cell-surface marker allows enrichment levels similar to that obtained with complex multicolor strategies. Altogether, our results indicate that even an extremely homogeneous HSC population, based on phenotype and dye efflux, cannot reconstitute mice at absolute efficiencies.