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  • 1
    ISSN: 1432-0878
    Keywords: Osteoblasts ; Preosteoclasts ; Cell differentiation ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Osteoblasts are involved in the bone resorption process by regulating osteoclast maturation and activity. In order to elucidate the mechanisms underlying osteoblast/preosteoclast cell interactions, we developed an in vitro model of co-cultured human clonal cell lines of osteoclast precursors (FLG 29.1) and osteoblastic cells (Saos-2), and evaluated the migratory, adhesive, cytochemical, morphological, and biochemical properties of the co-cultured cells. In Boyden chemotactic chambers, FLG 29.1 cells exhibited a marked migratory response toward the Saos-2 cells. Moreover, they preferentially adhered to the osteoblastic monolayer. Direct co-culture of the two cell types induced: (1) positive staining for tartrate-resistant acid phosphatase in FLG 29.1 cells; (2) a decrease of the alkaline phosphatase activity expressed by Saos-2 cells; (3) the appearance of typical ultrastructural features of mature osteoclasts in FLG 29.1 cells; (4) the release into the culture medium of granulocyte-macrophage colony stimulating factor. The addition of parathyroid hormone to the co-culture further potentiated the differentiation of the preosteoclasts, the cells tending to fuse into large multinucleated elements. These in vitro interactions between osteoblasts and osteoclast precursors offer a new model for studying the mechanisms that control osteoclastogenesis in bone tissue.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0878
    Keywords: Key words: Insulin-like growth factor-1 (IGF-I) ; Bone endothelial cells ; Preosteoclasts ; Co-culture ; Cell migration ; Electron microscopy (transmission and scanning) ; Bovine ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Little is known about the factors and the mechanisms involved in preosteoclast emigration from the vasculature. In this study, an in vitro model of bone endothelial lining was mimicked by culturing bone endothelial (BBE) cells at confluence on a 3-μm pore polycarbonate membranes. Preosteoclastic (FLG 29.1) cells were then added on top of the BBE cell monolayer and 10 nM insulin-like growth factor-1 (IGF-I) was added below the supporting membrane. Scanning and transmission electron microscopy were used to evaluate the chemotactic responses of preosteoclastic FLG 29.1 cells towards the IGF-I generated gradient. IGF-I potently stimulated chemotaxis in the FLG 29.1 cells, as shown by the migration of the preosteoclastic cells across the underlying BBE and through the intercellular junctions between adjacent endothelial cells. Subsequently, FLG 29.1 cells penetrated the pores of the supporting membrane and reached the lower face of the membrane. Thus, IGF-I, which is abundantly present in the bone tissue microenvironment, may play a paracrine role in the recruitment of the circulating preosteoclasts from the vascular compartment into the bone tissue. This in vitro model, which mimicks the in vivo phenomenon of preosteoclast extravasation, should prove useful in elucidating the molecular mechanisms that underlie this process.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-0878
    Keywords: Key words: Osteoblasts ; Preosteoclasts ; Cell differentiation ; Human
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. Osteoblasts are involved in the bone resorption process by regulating osteoclast maturation and activity. In order to elucidate the mechanisms underlying osteoblast/preosteoclast cell interactions, we developed an in vitro model of co-cultured human clonal cell lines of osteoclast precursors (FLG 29.1) and osteoblastic cells (Saos-2), and evaluated the migratory, adhesive, cytochemical, morphological, and biochemical properties of the co-cultured cells. In Boyden chemotactic chambers, FLG 29.1 cells exhibited a marked migratory response toward the Saos-2 cells. Moreover, they preferentially adhered to the osteoblastic monolayer. Direct co-culture of the two cell types induced: (1) positive staining for tartrate-resistant acid phosphatase in FLG 29.1 cells; (2) a decrease of the alkaline phosphatase activity expressed by Saos-2 cells; (3) the appearance of typical ultrastructural features of mature osteoclasts in FLG 29.1 cells; (4) the release into the culture medium of granulocyte-macrophage colony stimulating factor. The addition of parathyroid hormone to the co-culture further potentiated the differentiation of the preosteoclasts, the cells tending to fuse into large multinucleated elements. These in vitro interactions between osteoblasts and osteoclast precursors offer a new model for studying the mechanisms that control osteoclastogenesis in bone tissue.
    Type of Medium: Electronic Resource
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