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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1993 Mar;84(3):290–297. doi: 10.1111/j.1349-7006.1993.tb02869.x

Establishment and Characterization of a Simian Virus 40‐Immortalized Osteoblastic Cell Line from Normal Human Bone

Hideki Chiba 1,2,✉, Norimasa Sawada 1, Takao Ono 3, Seiichi Ishii 2, Michio Mori 1
PMCID: PMC5919145  PMID: 8387478

Abstract

We established a human osteoblastic cell line immortalized by simian virus 40 (SV40) in vitro, and designated it SV‐HFO. Immunocytochemically, the cells were positive for SV40 large T‐antigen, vimentin and osteocalcin, but negative for keratin and epithelial membrane antigen. The cells had characteristic morphologic and ultrastructural features of osteoblasts, produced alkaline phosphatase, and synthesized osteocalcin, the levels of which were elevated by treatment of the cells with 1α,25‐dihydroxyvitamin D3. The cells proliferated and showed such osteoblastic properties even under serum‐free conditions. The cells grew in soft agar, but did not form tumors when transplanted into athymic nude mice. Karyotypic analysis by the Q‐banding technique showed that these cells were of human origin. The SV‐HFO cell line is expected to serve as a suitable model for studying metabolism and carcinogenesis in human bone.

Keywords: Osteoblast, Human, Simian virus 40, Cell line, Osteocalcin

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REFERENCES

  • 1. ) Wong , G.Isolation and behavior of isolated bone‐forming cells . In “ Bone, Vol. 1: The Osteoblast and Osteocyte ,” ed. Hall B. K. , pp. 171 – 192 ( 1990. ). The Telford Press; , Caldwell . [Google Scholar]
  • 2. ) Kodama , H. , Amagai , Y. , Sudo , H. , Kasai , S. and Yamamoto , S.Establishment of a clonal osteogenic cell line from newborn mouse calvaria . Jpn. J. Oral Biol. , 23 , 899 – 901 ( 1981. ). [Google Scholar]
  • 3. ) Sudo , H. , Kodama , H. , Amagai , Y. , Yamamoto , S. and Kasai , S.In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria . J. Cell Biol. , 96 , 191 – 198 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4. ) Aubin , J. E. , Heersche , J. N. M. , Merriless , M. J. and Sodek , J.Isolation of bone cell clones with differences in growth, hormone responses, and extracellular matrix production . J. Cell Biol. , 92 , 452 – 461 ( 1982. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5. ) Grigoriadis , A. E. , Heersche , J. N. M. and Aubin , J. E.Differentiation of muscle, fat, cartilage, and bone from progenitor cells present in a bone‐derived clonal cell population: effect of dexamethasone . J. Cell Biol. , 106 , 2139 – 2151 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6. ) Heath , J. K. , Rodan , S. B. , Yoon , K. and Rodan , G. A.Rat calvarial cell lines immortalized with SV‐40 large T antigen: constitutive and retinoic acid‐inducible expression of osteoblastic features . Endocrinology , 124 , 3060 – 3068 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 7. ) Yamaguchi , A. and Kahn , A. J.Clonal osteogenic cell lines express myogenic and adipocytic developmental potential . Calcif. Tissue Int. , 49 , 221 – 225 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 8. ) Majeska , R. J. , Rodan , S. B. and Rodan , G. A.Parathyroid hormone‐responsive clonal cell lines from rat osteosarcoma . Endocrinology , 107 , 1494 – 1503 ( 1980. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Partridge , N. C. , Frampton , R. J. , Eisman , J. A. , Michelangeli , V. P. , Elms , E. , Bradley , T. R. and Martin , T. J.Receptors for 1,25(OH)2‐vitamin D3 enriched in cloned osteoblast‐like rat osteogenic sarcoma cells . FEBS Lett. , 115 , 139 – 142 ( 1980. ). [DOI] [PubMed] [Google Scholar]
  • 10. ) Partridge , N. C. , Alcorn , D. , Michelangeli , V. P. , Ryan , G. and Martin , T. J.Morphological and biochemical characterization of four clonal osteogenic sarcoma cell lines of rat origin . Cancer Res. , 43 , 4308 – 4314 ( 1983. ). [PubMed] [Google Scholar]
  • 11. ) Fogh , J. and Trempe , G.New human tumor cell lines . In “ Human Tumor Cell Lines In Vitro ,” ed. Fogh J. , pp. 115 – 159 ( 1975. ). Plenum Press; , New York . [Google Scholar]
  • 12. ) Franceschi , R. T. , James , W. M. and Zerlauth , G.1α,25‐Dihydroxyvitamin D3 specific regulation of growth, morphology and fibronectin in a human osteosarcoma cell line . J. Cell. Physiol. , 123 , 401 – 409 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 13. ) Rodan , S. G. , Imai , Y. , Thiede , M. A. , Wesolowski , G. , Thompson , D. , Bar‐Shavit , Z. , Shull , S. , Mann , K. and Rodan , G. A.Characterization of a human osteosarcoma cell line (SAOS‐2) with osteoblastic properties . Cancer Res. , 47 , 4961 – 4966 ( 1987. ). [PubMed] [Google Scholar]
  • 14. ) Fournier , B. and Price , P. A.Characterization of a new human osteosarcoma cell line OHS‐4 . J. Cell Biol. , 114 , 577 – 583 ( 1991. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15. ) Ohta , T. , Mori , M. , Koshiba , H. , Takada , J. , Matsuyama , T. and Ishii , S.Production of monoclonal antibodies specific for human bone 7‐carboxyglutamic acid containing protein . Virchows Arch. A Pathol. Anat. , 415 , 459 – 466 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 16. ) Lowry , O. H.Micromethods for the assay of enzyme II. Specific procedures. Alkaline phosphatase . Methods Enzymol. , 4 , 371 – 372 ( 1955. ). [Google Scholar]
  • 17. ) Macpherson , I. and Montagnier , L.Agar suspension culture for the selective assay of cells transformed by polyoma virus . Virology , 23 , 291 – 294 ( 1964. ). [DOI] [PubMed] [Google Scholar]
  • 18. ) Yoshida , M. C. , Ikeuchi , T. and Sasaki , M.Differential staining of parental chromosomes in interspecific cell hybrids with a combined quinacrine and 33258 Hoechst technique . Proc. Jpn. Acad. , 51 , 184 – 187 ( 1975. ). [Google Scholar]
  • 19. ) Sack , G. H. , Jr.Human cell transformation by simian virus 40, A review . In Vitro , 17 , 1 – 19 ( 1981. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Chang , S. E.In vitro transformation of human epithelial cells . Biochim. Biophys. Acta , 823 , 161 – 194 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 21. ) Miranda , A. F. , Babiss , L. E. and Fisher , P. B.Transformation of human skeletal muscle cells by simian virus 40 . Proc. Natl. Acad. Sci. USA , 80 , 6581 – 6585 ( 1983. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22. ) Steinberg , M. L. and Defendi , V.Altered pattern of growth and differentiation in human keratinocytes infected by simian virus 40 . Proc. Natl. Acad. Sci. USA , 76 , 801 – 805 ( 1979. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. ) Christian , B. J. , Loretz , L. J. , Oberley , T. D. and Reznikoff , C. A.Characterization of human uroepithelial cells immortalized in vitro by simian virus 40 . Cancer Res. , 47 , 6066 – 6073 ( 1987. ). [PubMed] [Google Scholar]
  • 24. ) Kaighn , M. E. , Narayan , K. S. , Ohnuki , Y. , Jones , L. W. and Lechner , J. F.Differential properties among clones of simian virus 40‐transformed human epithelial cells . Carcinogenesis , 1 , 635 – 645 ( 1980. ). [DOI] [PubMed] [Google Scholar]
  • 25. ) Yoon , K. , Rutledge , S. J. C. , Buenaga , R. F. and Rodan , G. A.Characterization of the rat osteocalcin gene: stimulation of promoter activity by 1,25‐dihydroxyvitamin D3 . Biochemistry , 27 , 8521 – 8526 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 26. ) Lian , J. , Stewart , C. , Puchacz , E. , Mackowiak , S. , Shalhoub , V. , Collart , D. , Zambetti , G. and Stein , G.Structure of the rat osteocalcin gene and regulation of vitamin D‐dependent expression . Proc. Natl. Acad. Sci. USA , 86 , 1143 – 1147 ( 1989. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27. ) Vermeulen , A. H. M. , Vermeer , C. and Bosnian , F. T.Histochemical detection of osteocalcin in normal and pathological human bone . J. Histochem. Cytochem. , 37 , 1503 – 1508 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 28. ) Price , P. A. and Baukol , S. A.1,25‐Dihydroxyvitamin D3 increases synthesis of the vitamin K‐dependent bone protein by osteosarcoma cells . J. Biol. Chem. , 255 , 11660 – 11663 ( 1980. ). [PubMed] [Google Scholar]
  • 29. ) Mukins , M. A. , Manolagas , S. C. , Deftos , L. J. and Sussman , H. H.1,25‐Dihydroxyvitamin D3 increases bone alkaline phosphatase isoenzyme levels in human osteogenic sarcoma cells . J. Biol. Chem. , 258 , 6219 – 6225 ( 1983. ). [PubMed] [Google Scholar]
  • 30. ) Franceschi , R. T. , Romano , P. R. and Park , K.‐Y.Regulation of type I collagen synthesis by 1,25‐dihydroxyvitamin D3 in human osteosarcoma cells . J. Biol. Chem. , 263 , 18938 – 18945 ( 1988. ). [PubMed] [Google Scholar]
  • 31. ) Spiess , Y. H. , Price , P. A. , Deftos , J. L. and Manolagas , S. C.Phenotype‐associated changes in the effects of 1,25‐dihydroxyvitamin D3 on alkaline phosphatase and bone Gla‐protein of rat osteoblastic cells . Endocrinology , 118 , 1340 – 1346 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 32. ) Lajeuness , D. , Frondoza , C. , Schoffield , B. and Sacktor , B.Osteocalcin secretion by the human osteosarcoma cell line MG‐63 . J. Bone Miner. Res. , 5 , 915 – 922 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 33. ) Barrett , J. C. and Fletcher , W. F.Cellular and molecular mechanisms of multistep carcinogenesis in cell culture models . In “ Mechanisms of Environmental Carcinogenesis, Vol. 2: Multistep Models of Carcinogenesis ,” ed. Barrett J. C. , pp. 73 – 116 ( 1987. ). CRC Press; , Boca Raton . [Google Scholar]

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