Summary
The osteogenic potential of the two human osteosarcoma cell lines HOS and KHOS; a cell line produced by the transformation of the HOS cells by the Kirsten murine sarcoma virus, was studied in vitro. HOS cells cultured more than 2 weeks formed nodules composed of two morphologically distinct layers, an epithelial-like surface cell layer and a collagen-rich inner cell layer. Alkaline phosphatase (ALPase) activity occurred in the plasma membrane of the surface cell layer, and calcified substances developing along collagen fibers were detected in the collagen-rich inner cell layer. The calcified substances were further examined by analytical electron microscopy and were shown to be hydroxyapatite crystals. In contrast, there was neither ALPase nor the deposition of a calcified substance in the KHOS cells.
Similar content being viewed by others
References
Alema S, Tato F, Boettiger D (1985) myc and src oncogenes have complementary effects on cell proliferation and expression of specific extracellular matrix components in definitive chondroblasts. Mol Cell Biol 5:538–544
Allebach ES, Boettiger D, Pacifici M, Adams SL (1985) Control of types I and collagen and fibronectin gene expression in chondrocytes delineated by viral transformation. Mol Cell Biol 5:1002–1008
Anderson HC (1985) Matrix vesicle calcification: Review and update. In: Peck WA (ed) Bone and mineral research, vol 3. Elsevier Science Publishers, Amsterdam, pp 109–149
Bar-Sagi D, Feramisco JR (1986) Induction of membrane ruffling and fluid-phase pinocytosis in quiescent fibroblast by ras proteins. Science 233:1061–1068
Bhargava U, Bar-Lev M, Bellows CG, Aubin JE (1988) Ultrastructural analysis of bone nodules formed in vitro by isolated fetal rat calvaria cells. Bone 9:155–163
Bonucci E (1978) Matrix vesicle formation in cartilage of scorbutic guinea pigs: Electron microscope study of serial sections. Metab Bone Dis Relat Res 1:205–212
Bonucci E, Silverstrini G (1984) Electron miceoscope investigation on the origin of matrix vesicles in bone. In: Cohn DV, Potts JT Jr, Fujita T (eds) Endocrine control of bone and calcium metabolism, vol 8B. Excerpta Medica, Amsterdam, pp 414–417
Boyan BD, Schwartz Z, Bonewald LF, Swain LD (1989) Localization of l,25-(OH72D3-responsive alkaline phosphatase in osteoblast-like cells (ROS 17/2.8, MG 63, and MC3T3) and growth cartilage cells in culture. J Biol Chem 264:11879–11886
Burstone MS (1958) Histochemical comparison of naphthol ASphosphate for the demonstration of phosphatases. J Natl Cancer Inst 20:601–615
Clemens TL, Garrett KP, Zhou XY, Pike JW, Harssler MR, Dempster DW (1988) Immunocytochemical localization of the 1,25-dihydroxyvitamin D3 receptor in target cells. Endocrinology 122:1224–1230
Colletta G, Pinto A, Di Fiore PP, Fusco AF, Ferrentino M, Avvedimento VE, Tsuchida N, Vecchio G (1983) Dissociation between transformed and differentiated phenotype in rat thyroid epithelial cells after transformation with a temperature-sensitive mutant of the Kirsten murine sarcoma virus. Mol Cell Biol 11:2099–2109
Dahl LK (1952) A simple and sensitive histochemical method for calcium. Proc Soc Exp Biol Med 80:474–479
Falcone G, Boettiger D, Alema S, Tato F (1984) Role of cell division in differentiation of myoblasts infected with a temperaturesensitive mutant of Rous sarcoma virus. EMBO J 3:1327–1331
Franceschi RT, James WM, Zerlauth G (1985) la, 25-dihydroxyvitamin D3 specific regulation of growth, morphology, and fibronectin in a human osteosarcoma cell line. J Cell Physiol 123:401–409
Franceschi RT, Linson CJ, Peter TC, Romano PR (1987) Regulation of cellular adhesion and fibronectin synthesis by 1α, 25-dihydroxyvitamin D3. J Biol Chem 262:4165–4171
Franceschi RT, Romano PR, Park KY (1988) Regulation of type I collagen synthesis by 1,25-dihydroxyvitamin D3 in human osteosarcoma cells. J Biol Chem 263:18938–18945
Fusco A, Pinto A, Tramontano D, Tajana G, Vecchio G, Tsuchida N (1983) Block in the expression of differentiation markers of rat thyroid cells by transformation with Kirsten murine sarcoma virus. Cancer Res 42:618–626
Hirai G (1959) A contribution to the ultrastructure of the dental enamel by electron microscopy and electron diffraction. Okajima Fol Anat Jap 34:125–157
Kirsten WH, Mayer LA (1967) Morphologic responses to a murine erythroblastosis virus. J Natl Cancer Inst 39:311–335
Lassar AB, Thayer MJ, Overell RW, Weintraub H (1989) Transformation by activated ras or fos prevents myogenesis by inhibiting expression of MyoD1. Cell 56:659–667
Lee WR, Laurie J, Townsend AL (1975) Fine structure of a radiation-induced osteogenic sarcoma. Cancer 36:1414–1425
Liau G, Yamada Y, Crombrugghe BD (1985) Coodinate regulation of the levels of type III and type I collagen mRNA in most but not all mouse fibroblasts. J Biol Chem 260:531–536
Mayahara H, Hirano H, Saito T, Ogawa K (1967) The new lead citrate method for the ultracytochemical demonstration of activity of non-specific alkaline phosphatase (Orthophosphoric monoester phosphohydrolase). Histochemie 11:88–96
McAllister RM, Gardner MB, Green AE, Nichols WW, Landing BH (1971) Cultivation in vitro of cells derived from a human osteosarcoma. Cancer 27:387–402
McSheehy PMJ, Chambers TJ (1986) Osteoblast-like cells in the presence of parathyroid hormone release soluble factor that stimulates osteoclastic bone resorption. Endocrinology 119:1645–1659
McSheehy PMJ, Chambers TJ (1987) 1,25-dihydroxyvitamin D3 stimulates rat osteoblastic cells to release a soluble factor that increases osteoblastic bone resorption. J Clin Invest 30:425–429
Muhrad A, Stein H, Bab A, Sela J (1978) Fine structure and enzymes of matrix vesicles in osteosarcoma: Possible occurrence of contractile proteins. Metab Bone Dis Relat Res 1:227–233
Mulkins MA, Manolagas SC, Deftos LJ, Sussman HH (1983) 1,25-dihydroxyvitamin D3 increases bone alkaline phosphatase isoenzyme levels in human osteogenic sarcoma cells. J Biol Chem 258:6219–6225
Murray E, Provvedini D, Curran D, Catherwood B, Sussman H, Manologas S (1987) Characterization of a human osteoblastic osteosarcoma cell line (SAOS-2) with high bone alkaline phosphatase activity. J Bone Miner Res 2:231–238
Myrdal SE, Auersperg N (1986) An agents produced by virustransformed cells cause unregulated ruffling in untransformed cells. J Cell Biol 102:1224–1229
Rhim JS, Cho HY, Huebner RJ (1975) Non-producer human cells induced by murine sarcoma virus. Int J Cancer 15:23–29
Rodan SB, Fischer MK, Egan JJ, Epstein PM, Rodan GA (1984) The effect of dexamethasone on parathyroid hormone stimulation of adenylate cyclase in ROS 17/2.8 cells. Endocrinology 115:951–958
Rodan SB, Imai Y, Thiede MA, Wesoloeski 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
Roseberry HH, Hastings AB, Morse JK (1931) X-ray analysis of bone and teeth. J Biol Chem 90:395–411
Shiina S, Mizuhira V, Amakawa T, Futaesaku Y (1970) An analysis of the histochemical procedure for sodium ion detection. J Histochem Cytochem 18:644–649
Tenenbaum HC, Heershe NM (1982) Differentiation of osteoblasts formation of mineralized bone in vitro. Calcif Tissue Int 34:76–79
Tsuchida N, Ryder T, Ohtsubo E (1982) Nucleotide sequence of the oncogene encoding the p21 transforming protein of Kirsten murine sarcoma virus. Science 217:937–939
Watson P, Kang YH, Falk MC (1989) Cytochemical properties of osteoblast cell membrane domains. J Histochem Cytochem 37:1235–1246
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ikeda, T., Futaesaku, Y. & Tsuchida, N. In vitro differentiation of the human osteosarcoma cell lines, HOS and KHOS. Virchows Archiv B Cell Pathol 62, 199–206 (1992). https://doi.org/10.1007/BF02899683
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02899683