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

doi:10.3969/j.issn.1006-7795.2014.05.009

Abstract

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

CLC Number:

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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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3969/j.issn.1006-7795.2014.05.009

https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2014/V35/I5/572

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