Receptor activator of nuclear factor kappa B ligand-induced mature osteoclasts

doi:10.3969/j.issn.2095-4344.2013.24.002

Abstract

Abstract:

BACKGROUND: Osteoclast as a terminal cell is difficult to obtain, and the shortage of mature osteoclast strains limits its application. Previous studies have shown that stromal cell culture or co-culture is commonly used to obtain the osteoclasts, and the receptor activator of nuclear factor κB ligand and macrophage colony-stimulating factor can be used in combination in order to obtain the mature osteoclasts.
OBJECTIVE: To observe the biological characteristics of mouse monocyte macrophage RAW264.7, and to analyze the feasibility of receptor activator of nuclear factor κB ligand-mediated differentiation of monocyte macrophage RAW264.7 into osteoclasts.
METHODS: The RAW264.7 cells were cultured, and then the RAW264.7 cells were induced with receptor activator of nuclear factor κB ligand for 7 days to observe the tartrate-resistant acid phosphatase staining results. The positive tartrate-resistant acid phosphatase staining and nucleus ≥ 3 were considered as the osteoclasts. Phalloidin staining was used to observe fibrous actin ring, and toluidine blue staining was used to observe the resorption pits on the surface of bovine bone slice.
RESULTS AND CONCLUSION: RAW264.7 cells could be induced into tartrate-resistant acid phosphatase-positive multinuclear cells with receptor activator of nuclear factor κB ligand, and formed fibrous actin rings; round or ellipse bone resorption pits were found in bone slice surface by microscope. RAW264.7 cells were considered as a good preosteoclast model that used for research of osteoclast differentiation. Receptor activator of nuclear factor κB ligand can induce the differentiation and maturation of RAW264.7 cells, reduce the application of macrophage colony-stimulating factor and make the training system simpler and easier to operate, and the induced cells have the high purity suitable for biological and biochemical studies of osteoclasts.

Key words: tissue construction, bone tissue construction, osteoclasts, receptor activator of nuclear factor kappa B, ligand, RAW264.7 cells, cell differentiation, induce, identification, fibrous actin, bone resorption, provincial grants-supported paper

CLC Number:

R318

Chen Guo-xian, Wang Guo-rong, Lin Zong-jin, Li Guo-shan, Guo Chun-xian, Luo Yuan-biao, . Receptor activator of nuclear factor kappa B ligand-induced mature osteoclasts[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(24): 4380-4385.

Figures/Tables 4

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