人破骨细胞的体外分离培养及与骨髓瘤细胞的相互作用

doi:10.3969/j.issn.1006-7795.2014.05.009

首都医科大学学报 ›› 2014, Vol. 35 ›› Issue (5): 572-576.doi: 10.3969/j.issn.1006-7795.2014.05.009

人破骨细胞的体外分离培养及与骨髓瘤细胞的相互作用

张建华, 董春霞, 任方刚, 覃艳红, 杨林花

山西医科大学第二医院血液科, 太原 030001

收稿日期:2014-08-25 出版日期:2014-10-21 发布日期:2014-10-20
通讯作者: 张建华 E-mail:jh20721@163.com

Isolation and culture of human osteoclasts from peripheral blood and the interaction between osteoclasts and myeloma cells

Zhang Jianhua, Dong Chunxia, Ren Fanggang, Tan Yanhong, Yang Linhua

Department of Hematology, the Second Hospital of ShanXi Medical University, Taiyuan 030001, China

Received:2014-08-25 Online:2014-10-21 Published:2014-10-20

摘要/Abstract

摘要：

目的研究体外分离培养人破骨细胞的可行性及与骨髓瘤细胞的相互作用。方法分离外周血单个核细胞（peripheral blood mononuclear cells，PBMC）体外培养形成破骨细胞（osteoclasts，OC），PBMC-OC与骨髓瘤（multiple myeloma， MM）细胞进行共培养。结果 PBMC在完全OC培养基中培养4 d后形成定型的破骨细胞前体（osteoclast precursor cell，pOC）；8226细胞可诱导pOC迁移并促进其向成熟的OC分化；pOC经完全OC培养基继续培养6~10 d后形成OC，在无OC培养基条件下，MM细胞仍可促进OC存活；OC明显促进MM细胞增生，抑制去血清培养诱导的MM细胞死亡。结论体外培养正常人PBMC-OC具有可行性，MM细胞促进OC成熟，OC促进骨髓瘤细胞生长与存活。

关键词: 破骨细胞, 体外培养, 骨髓瘤细胞

Abstract:

Objective To evaluate the feasibility of isolation and culture of osteoclasts (OCs) in vitro, and investigate the interaction between OCs and myeloma cells.Methods Human OCs were isolated from peripheral blood mononuclear cells (PBMCs) in vitro according to the previously described procedures. The interaction between PBMC-OCs and MM cells was determined by coculture. Results In PBMC culture, after 4 days of incubation with osteoclastic medium, the nonadherent cells were removed, leaving most of the remaining adherent cells expressed (tartrate-resistant acid phosphatase) TRAP, were considered committed pOCs; 8226 cells actively attracted precursor OCs (pOCs) and induced OCs formation. When cultured for an additional 6-10 days in OC medium, pOCs differentiated into large, multinucleared OCs. When OCs were cocultured with myeloma cells, apparently all OCs remained alive after 7 days without addition of sRANKL and macrophage-colony stimulating factor (M-CSF). PBMC-OCs stimulated the proliferation of myeloma cells in cocultures and rescued MM cells death in serum-depleted cultures.Conclusion PBMC-OCs from healthy volunteers promoted MM cells growth and survival, MM cells actively attracted pOCs and induced OCs formation.

Key words: osteoclasts, culture in vitro, myeloma cells

中图分类号:

张建华, 董春霞, 任方刚, 覃艳红, 杨林花. 人破骨细胞的体外分离培养及与骨髓瘤细胞的相互作用[J]. 首都医科大学学报, 2014, 35(5): 572-576.

Zhang Jianhua, Dong Chunxia, Ren Fanggang, Tan Yanhong, Yang Linhua. Isolation and culture of human osteoclasts from peripheral blood and the interaction between osteoclasts and myeloma cells[J]. Journal of Capital Medical University, 2014, 35(5): 572-576.

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人破骨细胞的体外分离培养及与骨髓瘤细胞的相互作用

Isolation and culture of human osteoclasts from peripheral blood and the interaction between osteoclasts and myeloma cells

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