Identification of multiple osteoclast precursor populations in murine bone marrow
Identification of multiple osteoclast precursor populations in murine bone marrow
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DOI：
10.1359/jbmr.051007
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发表时间：
2006-01-01
影响因子：
6.2
通讯作者：
Aguila, HL
中科院分区：
文献类型：
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作者：
Jacquin, C;Gran, DE;Aguila, HL
用一系列造血标记物对小鼠骨髓进行分离，以确定其破骨祖细胞群体的特征。我们发现，在总骨髓中早期的破骨细胞活性是由C111b(-/低)CD45R(-)CD3(-)CD115(高)组分所包含的一群细胞所概括的。我们通过分析其细胞表面标记进一步鉴定了小鼠骨髓中破骨细胞前体细胞群的表型。材料和方法：我们使用荧光激活细胞分选(FACS)来鉴定含有破骨细胞前体细胞的BM细胞亚群。我们根据多个标志物对骨髓进行分离，并在巨噬细胞集落刺激因子(M-CSF)和RANKL的作用下培养2-6天。结果：CD45R(-)、CD11b(-/low)群体重现了BM早期的破骨细胞活性。此外，尽管以前的实验表明，CD45R(+)群体中的破骨细胞活性丰富，但我们发现高纯度的CD45R(+)BM在体外不能分化为破骨细胞。我们还发现CD45R(-)CD11b(高)BM细胞是破骨细胞前体细胞的低效来源。然而，CD45R(-)CD11b(-/低)部分的细胞在M-CSF培养早期(24小时内)瞬时上调CD11b。最后，用CD115和CD117进一步分离骨髓表明，随着破骨细胞前体细胞的成熟，它们下调CD117，但仍保持CD115(+)。奇怪的是，从骨髓中分离的CD117(-)(CD115(高))细胞在体外具有低的破骨细胞活性。结论：我们提供了一种在M-CSF和RANKL作用下能够在体外分化为OCL的准确的小鼠BM亚群的精细分析。
Murine BM was fractionated using a series of hematopoietic markers to characterize its osteoclast progenitor populations. We found that the early osteoclastogenic activity in total BM was recapitulated by a population of cells contained within the C111b(-/low) CD45R(-)CD3(-)CD115(high) fraction.Introduction: Osteoclasts are of hematopoietic origin and they have been shown to share the same lineage as macrophages. We further characterized the phenotype of osteoclast progenitor populations in murine bone marrow (BM) by analyzing their cell surface markers.Materials and Methods: We used fluorescence-activated cell sorting (FACS) to identify the subsets of BM cells that contained osteoclast progenitors. We fractionated BM according to several markers and cultured the sorted populations for a period of 2-6 days with macrophage-colony stimulating factor (M-CSF) and RANKL. The numbers of multinucleated osteoclast-like cells (OCLs) that formed in the cultures were counted.Results: We found that the CD45R(-)CD11b(-/low) population recapitulated the early osteoclastogenic activity of total BM. In addition, although previous experiments indicated that osteoclastogenic activity was enriched within the CD45R(+) population, we found that highly purified CD45R(+) BM was incapable of differentiating into osteoclasts in vitro. We also found that CD45R(-)CD11b(high) BM cells were an inefficient source of osteoclast progenitors. However, CD11b was transiently upregulated by cells of the CD45R(-)CD11b(-/low) fraction early (within 24 h) during culture with M-CSF. Finally, further fractionation of BM using CD115 and CD117 showed that, as osteoclast precursor cells matured, they downregulate CD117 but remain CD115(+). Curiously, pure populations of CD117(-) (CD115 (high)) cells isolated fresh from BM have low osteoclastogenic activity in vitro.Conclusions: We provided a refined analysis of the precise subpopulations of murine BM that are capable of differentiating into OCLs in vitro when treated with M-CSF and RANKL.