Deep imaging of bone marrow shows non-dividing stem cells are mainly perisinusoidal.
Deep imaging of bone marrow shows non-dividing stem cells are mainly perisinusoidal.
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DOI：
10.1038/nature15250
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发表时间：
2015-10-01
期刊：
影响因子：
64.8
通讯作者：
Morrison SJ
中科院分区：
文献类型：
--
作者：
Acar M;Kocherlakota KS;Murphy MM;Peyer JG;Oguro H;Inra CN;Jaiyeola C;Zhao Z;Luby-Phelps K;Morrison SJ
造血干细胞（HSC）位于血管周围的小生境，但位置仍然存在争议。HSC是罕见的，并且很少可以在薄组织切片中或在活体成像时发现，使得难以全面定位分裂和非分裂HSC。我们发现α-catulinGFP/+仅在0.02%的骨髓造血细胞中表达，包括几乎所有的HSC。1/3.5的α-catulin-GFP+c-kit+细胞产生辐射小鼠的长期多谱系重建，表明α-catulin-GFP+c-kit+细胞含有具有与可用的最佳标记物相当的纯度的HSC。我们能够对骨髓进行光学清除以进行深度共聚焦成像，从而能够对数千个α-catulin-GFP+c-kit+细胞进行成像，并对大段骨髓进行数字重建。α-catulin-GFP+c-kit+细胞的分布表明，HSC在骨髓中央比在骨表面附近更常见，并且在相对于干骺端的骨干中更常见。几乎所有的HSC都与瘦素受体+和Cxcl 12高的小生境细胞接触。大约85%的HSC位于窦状血管的10μm范围内。大多数HSC远离小动脉、移行区血管和骨表面。Ki-67+分裂型HSC和Ki-67−非分裂型HSC也是如此。因此，分裂和非分裂的HSC主要位于窦周龛中，瘦素受体+Cxcl 12高细胞遍布骨髓。
Hematopoietic stem cells (HSCs) reside in a perivascular niche but the location remains controversial. HSCs are rare and few can be found in thin tissue sections or upon live imaging, making it difficult to comprehensively localize dividing and non-dividing HSCs. We discovered that α-catulinGFP/+ was expressed by only 0.02% of bone marrow hematopoietic cells, including virtually all HSCs. One in 3.5 α-catulin-GFP+c-kit+ cells gave long-term multilineage reconstitution of irradiated mice, indicating that α-catulin-GFP+c-kit+ cells contain HSCs with a purity comparable to the best markers available. We were able to optically clear the bone marrow to perform deep confocal imaging, making it possible to image thousands of α-catulin-GFP+c-kit+ cells and to digitally reconstruct large segments of bone marrow. The distribution of α-catulin-GFP+c-kit+ cells indicated that HSCs were more common in central marrow than near bone surfaces and in the diaphysis relative to the metaphysis. Nearly all HSCs contacted Leptin Receptor+ and Cxcl12high niche cells. Approximately 85% of HSCs were within 10μm of a sinusoidal blood vessel. Most HSCs were distant from arterioles, transition zone vessels, and bone surfaces. This was true of Ki-67+ dividing HSCs and Ki-67− non-dividing HSCs. Dividing and non-dividing HSCs thus reside mainly in perisinusoidal niches with Leptin Receptor+Cxcl12high cells throughout the bone marrow.