A Rapid Protocol for Direct Isolation of Osteoclast Lineage Cells from Mouse Bone Marrow

Lei Dang, Nanxi Li, Xiaohao Wu, Dijie Li, Zongkang Zhang, Bao Ting Zhang, Aiping Lyu, Lin Chen, Ge Zhang*, Jin Liu*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Osteoclast lineage cells (OLCs), including osteoclast precursors (OCPs) and mature osteoclasts (MOCs), participate in bone remodeling and mediate pathologic bone loss. Thus, it is essential to obtain OLCs for exploring their molecular features in both physiological and pathological conditions in vivo. However, the conventional protocols for obtaining OLCs ex vivo are not only time-consuming, but also unable to capture the cellular status of OLCs in vivo. In addition, the current antibody-based isolation approaches, such as fluorescence-/ magnetic-activated cell sorting, are not able to obtain pure osteoclasts because no unique surface antigen for osteoclasts has been identified. Here, we develop a rapid protocol for directly isolating OLCs from mouse bone marrow through magnetic-activated cell sorting (MACS). This protocol can rapidly enrich OCPs and MOCs, respectively, depending on the expression of the distinctive surface markers at their differentiation stages. It is optimized to isolate OLCs from four mice concurrently, of which sorting procedure could be completed within ~5 h.

Original languageEnglish
Article numbere4338
JournalBio-protocol
Volume12
Issue number5
DOIs
Publication statusPublished - 5 Mar 2022

Scopus Subject Areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)
  • Plant Science

User-Defined Keywords

  • Calcitonin receptor
  • Magnetic-activated cell sorting
  • Osteoclast lineage cells
  • Osteoclast-associated receptor
  • Osteoporosis

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