Analysis of histone 2B-GFP retention reveals slowly cycling hematopoietic stem cells.
Analysis of histone 2B-GFP retention reveals slowly cycling hematopoietic stem cells.
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DOI：
10.1038/nbt.1517
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发表时间：
2009-01
影响因子：
46.9
通讯作者：
Hock, Hanno
中科院分区：
文献类型：
--
作者：
Foudi, Adlen;Hochedlinger, Konrad;Van Buren, Denille;Schindler, Jeffrey W.;Jaenisch, Rudolf;Carey, Vincent;Hock, Hanno
造血干细胞（HSCs）基于其对针对快速增殖细胞的细胞毒损伤的抗性，被认为很少分裂，并且一直被假定会保留诸如核苷酸类似物5 - 溴脱氧尿苷（BrdU）这样的标记。然而，最近已经证明，BrdU保留对于造血干细胞既不敏感也不特异。在此我们表明，在小鼠中组蛋白2B（H2B） - 绿色荧光蛋白（GFP）融合蛋白的瞬时转基因表达能够更好地标记造血干细胞，并结合其他标记物可改进对其周转的分析。用一种高度严格的标记组合（L⁻K⁺S⁺CD48⁻CD150⁺）鉴定的造血干细胞中H2B - GFP稀释的数学模型揭示了其增殖速率中意想不到的异质性，并表明约20%的造血干细胞以极低的速率周转（每天≤0.8 - 1.8%）。对具有不同H2B - GFP水平的L⁻K⁺S⁺CD48⁻CD150⁺造血干细胞进行前瞻性分离和移植显示，较高的H2B - GFP标记保留与更强的长期重建造血潜能相关。
Hematopoietic stem cells (HSCs) are thought to divide infrequently based on their resistance to cytotoxic injury targeted at rapidly cycling cells and have been presumed to retain labels such as the nucleotide analogue 5-bromodeoxyuridine (BrdU). However, recently it has been demonstrated that BrdU-retention is neither sensitive nor specific for HSCs. Here we show that transient, transgenic expression of a Histone2B (H2B)-Green Fluorescent Protein (GFP) fusion protein in mice allows superior labeling of HSCs and permits improved analysis of their turnover in combination with other markers. Mathematical modeling of H2B-GFP dilution in HSCs, identified with a highly stringent marker combination (L−K+S+CD48−CD150+), revealed unexpected heterogeneity in their proliferation rates and suggests that ~ 20% of HSCs turn over at an extremely low rate (≤ 0.8–1.8% per day). Prospective isolation and transplantation of L−K+S+CD48−CD150+ HSCs with different H2B-GFP levels revealed that higher H2B-GFP label retention correlates with superior long-term repopulation potential.
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