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An assay system utilizing devitalized bone for assessment of differentiation of osteoclast progenitors
Author(s) -
Amano Shigeru,
Hanazawa Shigemasa,
Kawata Yasuhiro,
Ohta Kazuzo,
Kitami Hidekazu,
Kitano Shigeo
Publication year - 1992
Publication title -
journal of bone and mineral research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.882
H-Index - 241
eISSN - 1523-4681
pISSN - 0884-0431
DOI - 10.1002/jbmr.5650070312
Subject(s) - osteoclast , progenitor cell , chemistry , microbiology and biotechnology , biology , stem cell , in vitro , biochemistry
The present study provides a novel assay system to examine the differentiation of osteoclast progenitors on devitalized bone slices. We used the population of bone cells liberated enzymatically from 14‐day‐old mouse embryonal calvariae as a source of osteoclast progenitors. The analysis of differentiation of osteoclast progenitors into preosteoclasts and mature osteoclasts was assessed in terms of the formation of TRAP‐positive cells and pits or resorption lacunae, respectively, on devitalized bone slices. Osteoclasts having bone‐resorbing activity appeared when the calvarial cell population was cultured in the presence of 1 α ,25‐(OH) 2 D 3 on devitalized bone slices. The resorbing activity increased in a 1 α ,25‐(OH) 2 D 3 dose‐related manner. However, calcitonin, a potent inhibitor of differentiation and activation of osteoclast lineage cells, reduced the area of the resorption lacunae in a dose‐dependent fashion. The bone‐resorbing cells on the bone slices expressed an obvious ruffled border and clear zone, structures specific to mature osteoclasts. These results suggest that osteoclast progenitors in the mouse calvarial population examined differentiated into mature osteoclasts in the presence of 1 α ,25‐(OH) 2 D 3 on devitalized bone slices. Further, using this assay system we assessed the effect of some other osteotropic factors on the differentiation of osteoclast progenitors to mature osteoclasts. IL‐1, IL‐6, and PTH increased the formation of TRAP‐positive cells and pits and the area of resorption lacunae in a dose‐dependent fashion. However, prostaglandin E 2 was unable to induce the formation of resorption lacunae, although a significant appearance of TRAP‐positive cells was observed at a concentration of 200 ng/ml. In contrast, the stimulating effect of these osteotropic factors on the formation of resorption lacunae was inhibited significantly by indomethacin treatment (200 ng/ml). This novel assay system is thus useful for study of the differentiation of osteoclast progenitors to preosteoclasts and mature osteoclasts on devitalized bone slices and also for assessment of the effect of cytokines and hormones on the differentiation of osteoclast progenitors.