Nose Michinari, Yamazaki Hidetoshi, Hagino Hiroshi, Morio Yasuo, Hayashi Shin-Ichi, Teshima Ryota
Division of Orthopaedic Surgery, Department of Medicine of Sensory and Motor Organs, Faculty of Medicine, Tottori University, Yonago, Tottori 683-8503, Japan.
J Bone Miner Metab. 2009;27(1):57-65. doi: 10.1007/s00774-008-0011-0. Epub 2008 Dec 12.
Osteolytic disorders cause serious problems for quality of life with aging. Osteolysis is performed by osteoclasts of the hematopoietic lineage that share some characteristics with monocytes and macrophages. As osteoclast precursors (pOCs) are present in peripheral blood, their characterization in osteolytic diseases may help us to understand risk factors. Although essential factors for osteoclastogenesis have been reported, the effective induction from pOCs in human peripheral blood mononuclear cells (PBMCs) to mature osteoclasts in culture requires further improvement. The aim of this study was development of an efficient culture system for human osteoclastogenesis and providing a simple system for the enrichment of pOCs from PBMCs. We employed coculturing of human PBMCs with a mouse stromal cell line. Significant numbers of tartrate-resistant acid phosphatase-positive (TRAP(+)) multinucleated osteoclasts (MNCs), which could resorb dentine slices, were efficiently induced in this culture condition. pOCs were enriched in an anti-CD16 antibody column-passed anti-CD14 antibody-bound cell population isolated by magnetic cell sorting. We compared the percentage of the CD14(high) CD16(dull) cell population, which mainly contained pOCs in PBMCs, from age-matched patients with rheumatoid arthritis (RA) and osteoporosis (OP), but it was comparable. However, the mean number of TRAP(+) MNCs generated in cultures from PBMCs of RA was higher. In contrast, the frequency of pOCs in PBMCs from OP was relatively higher. These results suggest the characteristics of pOCs from RA and OP may be different, because single pOCs from OP gave rise to lower numbers of osteoclasts than those from RA.
溶骨性疾病随着年龄增长会给生活质量带来严重问题。骨溶解由造血谱系的破骨细胞执行，这些破骨细胞与单核细胞和巨噬细胞有一些共同特征。由于破骨细胞前体（pOCs）存在于外周血中，对其在溶骨性疾病中的特征进行研究可能有助于我们了解风险因素。尽管已经报道了破骨细胞生成的关键因素，但在培养中从人外周血单个核细胞（PBMCs）中的pOCs有效诱导生成成熟破骨细胞仍需进一步改进。本研究的目的是开发一种高效的人破骨细胞生成培养系统，并提供一种从PBMCs中富集pOCs的简单系统。我们采用人PBMCs与小鼠基质细胞系共培养。在这种培养条件下，可有效诱导出大量抗酒石酸酸性磷酸酶阳性（TRAP(+)）的多核破骨细胞（MNCs），这些细胞能够吸收牙本质切片。通过磁珠细胞分选法分离出抗CD16抗体柱通过的抗CD14抗体结合细胞群体，其中富含pOCs。我们比较了年龄匹配的类风湿关节炎（RA）和骨质疏松症（OP）患者外周血单个核细胞中主要包含pOCs的CD14(高)CD16(低)细胞群体的百分比，但两者相当。然而，RA患者PBMCs培养产生的TRAP(+)MNCs的平均数量更高。相比之下，OP患者PBMCs中pOCs的频率相对较高。这些结果表明，RA和OP患者的pOCs特征可能不同，因为来自OP的单个pOCs产生的破骨细胞数量比来自RA的少。
