High-dose PMA with RANKL and MCSF induces THP-1 cell differentiation into human functional osteoclasts in vitro
High-dose PMA with RANKL and MCSF induces THP-1 cell differentiation into human functional osteoclasts in vitro
复制标题
高剂量 PMA 与 RANKL 和 MCSF 诱导 THP-1 细胞体外分化为人功能性破骨细胞
DOI：
10.3892/mmr.2017.7625
复制
发表时间：
2017-12-01
影响因子：
3.4
通讯作者：
Zhang, Zhi Gang
中科院分区：
文献类型：
--
作者：
Li, Zhuo Hao;Si, Yu;Zhang, Zhi Gang
破骨细胞是一种大的多核细胞，在侵蚀性骨病中起作用。然而，不可能将破骨细胞从皮质骨中分离出来，以便培养细胞进行进一步的实验。因此，需要一个人类破骨细胞模型来研究骨破坏的潜在机制。最常用的破骨细胞模型是RAW264.7细胞系，一种小鼠单核巨噬细胞系；然而，目前还没有使用人类细胞系的可靠的破骨细胞模型。本研究的目的是利用THP-1细胞系建立功能性破骨细胞模型。用5或100 ng/ml佛波酯(PMA)刺激悬浮的THP-1细胞2天，诱导其分化为贴壁巨噬细胞。用50 ng/ml核因子-kappaB受体激活剂(RANKL)和巨噬细胞集落刺激因子(MCSF)刺激10d，诱导巨噬细胞向破骨细胞分化。大剂量PMA联合RANKL和MCSF处理后，THP-1细胞能够形成抗酒石酸酸性磷酸酶阳性破骨细胞，在骨吸收试验中能够吸收骨。小剂量PMA联合RANKL和MCSF未能诱导THP-1细胞向破骨细胞分化。PMA单独，或RANKL和MCSF单独组合，不足以刺激THP-1细胞分化为破骨细胞。在本研究中，使用THP-1细胞系建立了可靠的人破骨细胞模型。该破骨细胞模型可能为进一步研究提供有用的工具。
Osteoclasts are large multinuclear cells, which serve role in erosive bone disease. However, it is not possible to separate osteoclasts from cortical bone in order to culture the cells for further experiments. Therefore, a human osteoclast model is required to investigate the underlying mechanism of bone destruction. The most commonly-used osteoclast model is the RAW264.7 cell line, a murine mononuclear macrophage cell line; however, there exists no reliable osteoclast model using a human cell line. The aim of the present study was to establish a functional osteoclast model using the THP-1 cell line. Suspended THP-1 cells were stimulated for 2 days with 5 or 100 ng/ml phorbol-12 myristate-13 acetate (PMA) in order to induce the cells to differentiate into adherent macrophages. A 10-day stimulation with 50 ng/ml receptor activator of nuclear factor kappa-B ligand (RANKL) and macrophage colony-stimulating factor (MCSF) was performed in order to induce macrophage differentiation into osteoclasts. Treatment with high-dose PMA with RANKL and MCSF enabled the THP-1 cells to form tartrate-resistant acid phosphatase-positive osteoclasts, which were able absorb bone in a bone resorption test. Treatment with low-dose PMA with RANKL and MCSF failed to induce THP-1 cell differentiation into osteoclasts. PMA alone, or a combination of RANKL and MCSF alone, is insufficient to stimulate THP-1 cell differentiation into osteoclasts. In the present study, a reliable human osteoclast model was established using the THP-1 cell line. This osteoclast model may provide a useful tool for further studies.