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.
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Akatsu T, Takahashi N, Udagawa N, Sato K, Nagata N, Moseley JM, Martin TJ, Suda T (1989) Parathyroid hormone (PTH)-related protein is a potent stimulator of osteoclast-like multinucleated cell formation to the same extend as PTH in mouse marrow cultures. Endocrinology 125:20–27
Baron R, Neff L, Tran Van P, Nefussi JR, Vignery A (1986) Kinetic and cytochemical identification of osteoclast precursors and their differentiation into multinucleated osteoclasts. Am J Pathol 122:363–378
Bleiberg I, Glowacki J, Anklesaria P, Greenberger JS (1992) Origin of stromal cells associated with osteoclast recruitment in s.c. implants of bone particles in chimeric mice. Exp Hematol 20:957–961
Boyden S (1962) The chemotactic effect of a mixture of antibody and antigen on polymorphonuclear leukocytes. J Exp Med 115:453–466
Braidman IP, Rothwell C, Webber DM, Crowe P, Anderson DC (1990) Location of osteoclast precursors in fetal rat calvaria cultured on collagen gel. J Bone Miner Res 5:287–298
Burger EH, Van Der Meer JWM, Nijweide PJ (1984) Osteoclast formation from mononuclear phagocytes: role of bone-forming cells. J Cell Biol 99:1901–1906
Canalis E (1983) Effect of glucocorticoids on type I collagen synthesis, alkaline phosphatase activity, and deoxyribonucleic acid content in cultured rat calvariae. Endocrinology 112:931–939
Chambers TJ, Fuller K (1985) Bone cells predispose bone surfaces to resorption by exposure of mineral to osteoclastic contact. J Cell Sci 76:155–165
Clark SC, Kamen R (1987) The human hematopoietic colonystimulating factors. Science 236:1229–1237
Coccia PF, Krivit W, Cervenka J, Clawson C, Kersey JK, Kim TH Nesbit ME, Ramsay NKC, Warkentin PI, Teitelbaum SL, Kahn AJ, Brown DM (1980) Successful bone-marrow transplantation for infantile malignant osteopetrosis. N Engl J Med 302:701–708
Cooper RA, Braunwald AD, Kuo AL (1982) Phorbol ester induction of leukemic cell differentiation is a membrane-mediated process. Proc Natl Acad Sci USA 79:2865–2869
Dedhar S, Gaboury L, Galloway P, Eaves C (1988) Human granulocyte-macrophage colony stimulating factor is a growth factor active on a variety of cell types of nonhemopoietic origin. Proc Natl Acad Sci USA 85:9253–9257
Dickson IR, Scheven BAA (1989) Regulation of new osteoclast formation by a bone cell-derived macromolecular factor. Biochem Biophys Res Commun 159:1383–1390
Ejiri S (1983) The preosteoclast and its cytodifferentiation into the osteoclast: ultrastructural and histochemical studies of rat fetal parietal bone. Arch Histol Jpn 46:533–556
Felix, Cecchini MG, Hofstetter W, Guenther HL, Fleisch H (1991) Production of granulocyte-macrophage (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) by rat clonal osteoblastic cell population CRP10/30-mycl stimulated by tumor necrosis factor. Endocrinology 126:661–667
Formigli L, Benvenuti S, Calabresi E, Paoletti C, Masi L, Bernabei PA, Brandi ML (1991) Differentiation of osteoclast precursors induced by osteoblastic cells. J Bone Miner Res 6:S94
Formigli L, Zecchi Orlandini S, Benvenuti S, Masi L, Pinto A, Gattei V, Bernabei PA, Gehron Robey P, Collin-Osdoby, Brandi ML (1995) “In vitro” structural and functional relationships between preosteoclastic and bone endothelial cells: a juxtacrine model for migration and adhesion of osteoclast precursors. J Cell Physiol (in press)
Fuller K Chambers TJ (1989) Bone matrix stimulates osteoclastic differentiation in cultures of rabbit bone-marrow cells. J Bone Miner Res 4:179–183
Fuller K, Gallagher AC, Chambers TJ (1991) Osteoclast resorption-stimulating activity is associated with the osteoblast cell surface and/or the extracellular matrix. Biochem Biophys Res Commun 181:67–73
Galvin RJS, Osdoby P (1991) Chick “osteoclast like” cell conditioned media inhibits osteoblast collagen synthesis and alkaline phosphatase activity. J Bone Miner Res 6[Suppl 1]:S143
Gattei V, Bernabei PA, Pinto A, Bezzini R, Ringressi A, Formigli L, Tanini A, Attadia V, Brandi ML (1992) Phorbol ester induced osteoclast-like differentiation of a novel human leukemic cell line (FLG 29.1). J Cell Biol 116:437–447
Gehron Robey P (1989) The biochemistry of bone. In Endocrinol Metab Clin North Am 18:859–897
Greenfield EM, Alvarez JI, McLaurine EA, Ousler MJ, Blair HC, Osdoby P, Teitelbaum SL, Ross FP (1992) Avian osteoblast conditioned media stimulate bone resorption by targeting multinucleated osteoclast precursors. Calcif Tissue Int 51:317–323
Hattersley G, Chambers TJ (1989) Generation of osteoclastic function in mouse bone-marrow cultures: multinuclearity and tartrate-resistant acid phosphatase are unreliable markers for osteoclastic differentiation. Endocrinology 124:1689–1696
Hiura K, Sumitani K, Kawata T, Higashino K, Okawa M, Sato T, Hakeda Y, Kumegawa M (1991) Mouse osteoblastic cells (MC3T3-E1) at different stages of differentiation have opposite effects on osteoclastic cells formation. Endocrinology 128:1630–1637
Horowitz MC, Coleman DL, Ryaby JT, Einhorn TA (1989) Osteotropic agents induce the differential secretion of granulocytemacrophage colony stimulating factor by the osteoblast cell line MC3T3-E1. J Bone Miner Res 4:911–921
Ibbotson KJ, Roodman GD, McManus LM, Mundy GR (1984) Identification and characterization of osteoclast-like cells and their progenitors in cultures of feline marrow mononuclear cells. J Cell Biol 99:471–480
Irie K, Ozawa H (1990) Relationship between tooth eruption, occlusion and alveolar bone resorption: histochemical and cytological studies of bone remodelling on rat incisor alveolar bone facing the enamel after root resection. Arch Histol Cytol 53:511–522
Kurihara N, Chenu C, Miller M, Civin C, Rodan GD (1990) Identification of committed mononuclear precursors for osteoclast-like cells formed in long term human marrow cultures. Endocrinology 126:2733–2741
Luk SC, Nopajaroonsri C, Simon GT (1974) The ultrastructure of endosteum: a topographic study in young adult rabbits. J Ultrastruct Res 46:165–183
Malone JD, Teitelbaum SL, Griffin GL, Senior RM, Kahn AJ (1982) Recruitment of osteoclast precursors by purified bone matrix constituents. J Cell Biol 92:227–230
Marks SC (1983) The origin of osteoclasts. J Oral Pathol 12: 226–256
Marshall MJ, Nisbet NW, Green PM (1986) Evidence for osteoclast production in mixed bone cell cultures. Calcif Tissue Int 38:268–274
McDonald BR, Takahashi N, McManus LM, Holahan J, Mundy GR, Roodman GD (1987) Formation of multinucleated cells that respond to osteotropic hormones in long term human bone marrow cultures. Endocrinology 120:2326–2333
McSheehy PMJ, Chambers TJ (1986a) Osteoblastic cells mediate osteoclastic responsiveness to parathyroid hormone. Endocrinology 118:824–828
McSheehy PMJ, Chambers TJ (1986b) Osteoblast-like cells in the presence of parathyroid hormone release soluble factor that stimulates osteoclastic bone resorption. Endocrinology 119: 1654–1659
Metcalf D (1986) The molecular biology and function of the granulocyte-macrophage colony stimulating factors. Blood 67: 257–267
Mochizuki H, Hakeda Y, Wakatsuki N, Usui N Akashi S, Sato T, Tanaka K, Kumegawa M (1992) Insulin-like growth factor I supports formation and activation of osteoclasts. Endocrinology 131:1075–1080
Osdoby P, Oursler MJ, Salino-Hugg T, Krukowski M (1988) Osteoclast development: the cell surface and the bone enviroment. Ciba Found Symp 136:108–124
Pegoraro L, Abrham JL, Cooper RA, Levis A, Lange B, Meo P, Rovera G (1980) Differentiation of human leukemias in response to 12-O-tetradecanoylphorbol-13-acetate in vitro. Blood 55:859–862
Perry HM, Gurbani S (1992) Development of monoclonal antibodies to parathyroid hormone-induced resorptive factors from osteoblast-like cells. Calcif Tissue Int 50:237–244
Rodan SB, Imai Y, Thiede MA, Wesolowski G, Thompson D, Bar-Shavit Z, Shull S, Mann K, Rodan GA (1987) Characterization of a human osteosarcoma cell line (Saos-2) with osteoblastic properties. Cancer Res 47:4961–4966
Streeten E, Ornberg R, Curcio F, Sakaguchi K, Marx S, Auerbach GB, Brandi ML (1989) Cloned endothelial cells from fetal bovine bone. Proc Natl Acad Sci USA 86:916–920
Takahashi N, Akatsu T, Udagawa N, Sasaki T, Yamaguchi A, Moseley JM, Martin JT, Suda T (1988a) Osteoblastic cells are involved in osteoclast formation. Endocrinology 123:2600–2602
Takahashi N, Yamana H, Yoshiki S, Roodman GD, Mundy GR, Jones SJ, Boyde A, Suda T (1988b) Osteoclast-like cell formation and its regulation by osteotropic hormones in mouse bone marrow cultures. Endocrinology 122:1373–1382
Takahashi N, Udagawa N, Akatsu T, Tanaka H, Shionome M, Suda T (1991) Role of colony-stimulating factors in osteoclast development. J Bone Miner Res 6:977–985
Teti A, Grano M, Colucci S, Cantatore FP, Loperfido MC, Zambonin Zallone A (1991) Osteoblast-osteoclast relationships in bone resorption: osteoblasts enhance osteoclast activity in a serum-free co-culture system. Biochem Biophys Res Commun 179:634–640
Thomson BM, Saklatvala J, Chambers TJ (1986) Osteoblasts mediate interleukin I stimulation of bone resorption by rat osteoclasts. J Exp Med 164:104–112
Tran Van P, Vignery A, Baron R (1982) An electron-microscopic study of bone-remodeling sequence in the rat. Cell Tissue Res 225:283–292
Van De Wijngaert FP, Schipper CA, Tas MC, Burger EH (1988) Role of mineralizing cartilage in osteoclast and osteoblast recruitment. Bone 9:81–88
Walker DG (1972) Congenital osteopetrosis in mice cured by parabiotic union with normal siblings. Endocrinology 91:916–920
Webber DM, Menton D, Osdoby P (1990) An in vivo model system for the study of avian osteoclast recruitment and activity. Bone Miner 11:127–140
Yamaga M, Iwaku M, Ozawa H (1992) Alveolar bone remodelling in the early stage of experimental apical periodonitis in the rat mandible. Arch Histol Cytol 55:137–150
Yamashita T, Asano K, Takahashi N, Akatsu T, Udagawa N, Sasaki T, Martin TJ, Suda T (1990). Cloning of an osteoblastic cell line involved in the formation of osteoclast-like cells. J Cell Physiol 145:587–595
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Orlandini, S.Z., Formigli, L., Benvenuti, S. et al. Functional and structural interactions between osteoblastic and preosteoclastic cells in vitro. Cell Tissue Res 281, 33–42 (1995). https://doi.org/10.1007/BF00307956
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DOI: https://doi.org/10.1007/BF00307956