A Novel Osteoclast Precursor Cell Line, 4B12, Recapitulates the Features of Primary Osteoclast Differentiation an Function: Enhanced Transfection Efficiency Before and After Differentiation
A Novel Osteoclast Precursor Cell Line, 4B12, Recapitulates the Features of Primary Osteoclast Differentiation an Function: Enhanced Transfection Efficiency Before and After Differentiation
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DOI：
10.1002/jcp.21827
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发表时间：
2009-10-01
影响因子：
5.6
通讯作者：
Ohmori, Yoshihiro
中科院分区：
文献类型：
--
作者：
Amano, Shigeru;Sekine, Keisuke;Ohmori, Yoshihiro
破骨细胞是由单核细胞/巨噬细胞谱系前体分化而来的吸收骨的多核细胞。采用免疫荧光和细胞分选方法，从14日龄小鼠胚胎颅骨中提取mac - 1 (+)c-Fms(+)RANK（+）细胞，建立了一种新的破骨细胞前体细胞4B12。与M-CSF依赖性骨髓巨噬细胞（M-BMMs）一样，在RANKL存在下，4B12细胞需要M-CSF才能分化为TRAP阳性的多核细胞[TRAP(+) MNCs]。在牙本质切片上，从4B12细胞分化而来的骨吸收破骨细胞具有清晰带和褶皱边界，并表达破骨细胞特异性基因。il - 1 α存在时，骨吸收活性增强，而TRAP不增强。4B12细胞在牙本质切片上形成的TRAP(+) MNCs数量和凹坑数量是M-BMMs的4倍。4B12细胞被鉴定为巨噬细胞，具有mac - 1和F4/80标记，但在共聚焦激光扫描显微镜下发现，这些标记在分化为破骨细胞后失去了。4B12细胞不具有分化为树突状细胞的潜力，这表明了破骨细胞谱系的承诺。4B12细胞在分化前后均可通过siRNA转染。这些数据表明，4B12细胞忠实地复制了原代细胞的特性，是分析破骨细胞发生和破骨细胞功能的分子和细胞调控机制的有用和强大的模型。j .细胞。中国生物医学工程学报，21(2):444 - 444,2009。(C) 2009 Wiley-Liss, Inc。
Osteoclasts are bone-resorbing multinucleated cells differentiated from monocyte/macrophage lineage precursors. A novel osteoclast precursor cell line, 4B12 was established from Mac-I(+)c-Fms(+)RANK(+) cells from calvaria of 14-day-old mouse embryos using immunofluorescence and cell-sorting methods. Like M-CSF-dependent bone marrow macrophages (M-BMMs), M-CSF is required for 4B12 cells to differentiate into TRAP-positive multinucleated cells [TRAP(+) MNCs] in the presence of RANKL. Bone-resorbing osteoclasts differentiated from 4B12 cells on dentine slices possess both a clear zone and ruffled borders and express osteoclast-specific genes. Bone-resorbing activity, but not TRAP, was enhanced in the presence of IL-I alpha. The number of TRAP(+) MNCs and the number of pits formed from 4B12 cells on dentine slices was fourfold higher than that from M-BMMs. 4B12 cells were identified as macrophages with Mac-I and F4/80, yet lost these markers upon differentiation into osteoclasts as determined by confocal laser scanning microscopy. The 4B12 cells do not have the potential to differentiate into dendritic cells indicating commitment to the osteoclast lineage. 4B12 cells are readily transfectable with siRNA transfection before and after differentiation. These data show that 4B12 cells faithfully replicate the properties of primary cells and are a useful and powerful model for analyzing the molecular and cellular regulatory mechanisms of osteoclastogenesis and osteoclast function. J. Cell. Physiol. 221: 40-53, 2009. (C) 2009 Wiley-Liss, Inc.