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Updated: Jan 23, 2026

A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation
Published on: March 15, 2018
Cellular study of the LPS-induced osteoclastic multinucleated cell formation from RAW264.7 cells
Yuan Xiao1, Yang Cao1, Chengchao Song2,3
1Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong Univerisity, School of Medicine, Xi'an, Shaanxi, China.
Abstract:
Despite the response to the receptor activator of nuclear factor κ-Β ligand (RANKL), a study has reported that lipopolysaccharide (LPS) could induce RAW264.7 linage osteoclastic differentiation. However, on the contrary, another study recently showed that the LPS-induced multinuclear cells from RAW264.7 did not express osteoclastic functions. Interestingly, in our previous study, we found that RAW264.7 cells pretreated with 10 ng LPS plus macrophage-colony stimulating factor did not show any effects for enhancing RANKL-induced osteoclastic cell differentiation. Therefore, in our current study, we aim to investigate the oteoclastogesis induction ability and efficacy of LPS in the RAW264.7 cell line and relevant molecular signaling. The osteoclastogenic activity of LPS-treated RAW264.7 linage was studied by bone resorption pits and fibrous actin study. Besides that, through polymerase chain reaction and western blot, we showed that the transcriptional factor c-Fos and Nfatc1 might be associated with LPS-induced osteoclastogenesis. Overall, the results of our current study showed positive proof for osteoclast generation from LPS-independent treatment, as well as established an optimal and efficient method for this process.
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