Isolation and characterization of murine clonogenic osteoclast progenitors by cell surface phenotype analysis
Isolation and characterization of murine clonogenic osteoclast progenitors by cell surface phenotype analysis
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DOI：
10.1182/blood.v91.4.1272
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发表时间：
1998-02-15
期刊：
影响因子：
20.3
通讯作者：
Lee, MY
中科院分区：
文献类型：
--
作者：
Muguruma, Y;Lee, MY
破骨细胞是来源于造血的骨吸收细胞；然而，产生成熟破骨细胞的祖细胞群仍然难以捉摸。我们描述了克隆性破骨细胞祖细胞（集落形成单位-破骨细胞[CFU-O]）的独特细胞表面表型，并获得了选择性富集这些祖细胞的骨髓细胞群。根据物理和细胞表面特征依次分离全骨髓细胞，并检测CFU-O和其他造血祖细胞的存在。CFU-O在非粘附、低密度、谱系标记阴性（Lin(-)）中富集。Thy1.2阴性（Thy1.2(-)）， Sca1阴性（Sca1(-)）和c-kit阳性（c-kit(+)）人群，以及对巨噬细胞集落刺激因子（CSF; CFU-M），粒细胞-巨噬细胞-CSF （CFU-GM）和干细胞因子（CFU-SCF）有反应的祖细胞。根据c-kit荧光将Lin(-)Thy1.2(-)Sca1（-）群体分为c-kit（高）和c-kit（低）群体，在c.kit（高）群体中发现超过88%的CFU-M、CFU-GM和CFU-SCF。与上述造血祖细胞相比，CFU-O在c-kit（低）人群中明显更高：c-kit（低）人群中80%的祖细胞是CFU-O。c-kit（高）和c-kit（低）人群的CFU-O均表现出破骨细胞的关键特征：多核酒石酸抗性酸性磷酸酶阳性细胞形成，玻璃体粘连素受体、c-src和降钙素受体的表达以及骨吸收。我们已经在破骨细胞谱系的最早阶段确定了祖细胞群，并开发了一种将其从其他造血祖细胞中分离出来的方法。这将有助于为理解破骨细胞分化的分子机制铺平道路。(C) 1998年由美国血液病学会出版。
Osteoclasts are bone resorbing cells of hematopoietic origin; however, a progenitor cell population that gives rise to mature osteoclasts remains elusive. We have characterized a unique cell surface phenotype of clonogenic osteoclast progenitors (colony-forming unit-osteoclast [CFU-O]) and obtained a marrow cell population selectively enriched for these progenitors. Whole bone marrow cells were sequentially separated based on physical and cell surface characteristics, and the presence of CFU-O and other hematopoietic progenitors was examined. CFU-O was enriched in a nonadherent, low-density, lineage-marker-negative (Lin(-)). Thy1.2-negative (Thy1.2(-)), Sca1-negative (Sca1(-)), and c-kit-positive (c-kit(+)) population, as were the progenitors that were responsive to macrophage-colony-stimulating factor(CSF; CFU-M), granulocyte-macrophage-CSF (CFU-GM), and stem cell factor (CFU-SCF). When the Lin(-)Thy1.2(-)Sca1(-) population was divided into c-kit(high) and c-kit(low) populations based on c-kit fluorescence, over 88% of CFU-M, CFU-GM, and CFU-SCF were found in the c.kit(high) population. In relation to the above mentioned hematopoietic progenitors, CFU-O was significantly higher in the c-kit(low) population: 80% of progenitors present in the c-kit(low) population were CFU-O. The CFU-O in both c-kit(high) and c-kit(low) populations showed key features of the osteoclast: multinucleated tartrate-resistant acid phosphatase-positive cell formation, expressions of vitronectin receptors, c-src and calcitonin receptors, and bone resorption. We have identified a progenitor cell population in the earliest stage of the osteoclast lineage so far described and developed a method to isolate it from other hematopoietic progenitors. This should help pave the way to understand the molecular mechanisms of osteoclast differentiation. (C) 1998 by The American Society of Hematology.