Evaluation of efficacy on RANKL induced osteoclast from RAW264.7 cells

Chengchao Song, Xiaobin Yang, Yongsheng Lei, Zhen Zhang, Wanli Smith, Jinglong Yan, Lingbo Kong

Research output: Contribution to journalArticle

Abstract

Established RAW264.7 cell lines for osteoclastic differentiation has been widely engaged in bone homeostasis research, however, the efficacy of RANKL independently stimulating has rarely been defined, because protocols were usually developed and modified by various laboratories. Otherwise, problematic issues are also lie in the cell's seeding density, RANKL stimulating time point, and distinguishing osteoclastogenesis ability of RANKL-treated RAW264.7 cells. Therefore, in the current study, we examined the efficacy of various concentrations of RANKL-treated RAW264.7 for its osteoclastic differentiation with or without pretreated other costimulators such as: LPS and/or M-CSF. The oteoclastogenesis ability of RANKL-treated RAW264.7 cells was demonstrated by bone resorption pit, F-actin, and osteoclastogenesis specific marker studies. Besides that, through tartrate-resistant acid phosphatase (TRAP) staining, we clarified to start the treatment with 30 ng/ml RANKL at 12 hr after seeded RAW264.7 with the density of 6.25 × 10 3 cells/cm 2 manifested an significantly increased number of multinucleated osteoclastic cells. Overall, our results establishing an optimal method for RANKL independently inducing RAW 264.7 cell osteoclastic differentiation, which could efficiently generate osteoclasts in vitro for significant advances in our understanding of bone biology.

Original languageEnglish (US)
JournalJournal of Cellular Physiology
DOIs
StateAccepted/In press - Jan 1 2018

Fingerprint

Osteoclasts
Bone
Osteogenesis
Macrophage Colony-Stimulating Factor
Acid Phosphatase
Bone and Bones
Actins
Bone Resorption
Cells
Cell Differentiation
Homeostasis
Cell Count
Staining and Labeling
Cell Line
Research

Keywords

  • bone marrow macrophages
  • osteoclast
  • osteoclastogenesis
  • protocol
  • RANKL
  • RAW264.7

ASJC Scopus subject areas

  • Physiology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Evaluation of efficacy on RANKL induced osteoclast from RAW264.7 cells. / Song, Chengchao; Yang, Xiaobin; Lei, Yongsheng; Zhang, Zhen; Smith, Wanli; Yan, Jinglong; Kong, Lingbo.

In: Journal of Cellular Physiology, 01.01.2018.

Research output: Contribution to journalArticle

Song, Chengchao ; Yang, Xiaobin ; Lei, Yongsheng ; Zhang, Zhen ; Smith, Wanli ; Yan, Jinglong ; Kong, Lingbo. / Evaluation of efficacy on RANKL induced osteoclast from RAW264.7 cells. In: Journal of Cellular Physiology. 2018.
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