Int J Biochem Mol Biol 2011;2(2):183-189
Original Article
A simplified method for the generation of human osteoclasts in vitro

James J Cody, Angel A Rivera, Jianzhong Liu, Jillian M. Liu, Joanne T Douglas, Xu Feng

Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, USA; Division of Human Gene Therapy, Departments of
Medicine, Obstetrics & Gynecology, Pathology and Surgery, University of Alabama at Birmingham, Birmingham, AL, USA; Division of Cardiology,
Department of Medicine, Emory University, Atlanta, GA, USA; The Gene Therapy Center, University of Alabama at Birmingham, Birmingham, AL,

Received April 18, 2011; accepted April, 2011; Epub April, 2011; Published May 15, 2011

Abstract: Osteoclasts are large, multinucleated cells responsible for the resorption of mineralized bone matrix.  These cells are critical players
in the bone turnover involved in bone homeostasis.  Osteoclast activity is connected to the establishment and expansion of skeletal
metastases from a number of primary neoplasms.  Thus, the formation and activation of osteoclasts is an area of research with many potential
avenues for clinical translation.  Past studies of osteoclast biology have utilized primary murine cells cultured in vitro.  Recently, techniques
have been described that involve the generation of osteoclasts from human precursor cells.  However, these protocols are often time-
consuming and insufficient for generating large numbers of osteoclasts.  We therefore developed a simplified protocol by which human
osteoclasts may be easily and reliably generated in large numbers in vitro. In this study, osteoclasts were differentiated from bone marrow
cells that had been aliquotted and frozen.  Cells were generated by culture with recombinant macrophage colony-stimulating factor (M-CSF)
and receptor activator of NF-κB ligand (RANKL).  Both human and murine RANKL were shown to efficiently generate osteoclasts, although
higher concentrations of murine RANKL were required.  Formation of osteoclasts was demonstrated qualitatively by tartrate-resistant acid
phosphatase (TRAP) staining.  These cells were fully functional, as confirmed by their ability to form resorption pits on cortical bone slices.
Functional human osteoclasts can be difficult to generate in vitro by current protocols.  We have demonstrated a simplified system for the
generation of human osteoclasts in vitro that allows for large numbers of osteoclasts to be obtained from a single donor. (IJBMB1104004).

Keywords: Osteoclast, protocol, bone resorption, human, bone marrow, in vitro

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Address all correspondence to:
Xu Feng, PhD
Department of Pathology
University of Alabama at Birmingham
1670 University Boulevard, VH G046
Birmingham, AL 35294-0019
Tel: 205-975-0990
Fax: 205-934-1775