Culture of cells of the osteoblast lineage

  • Francis J. Hughes
  • Jane E. Aubin


Osteoblastic cell cultures have been utilized to study a wide range of issues in bone biology, including the regulation of the differentiation and metabolic activity of bone cells. These studies have included, for example: the investigation of the action of hormones, cytokines and other signalling molecules on bone cells; the molecular mechanisms of action of such factors; studies of the osteoblast cell lineage and differentiation-dependent changes in phenotype; the synthesis of matrix proteins and other secreted molecules; and studies of the interaction of bone cells with biomaterials. Some obvious advantages of such methods over experiments conducted in vivo include the ability to control more fully the cells being assessed and their environment, the relative ease of sampling and analysing changes in the parameters under investigation, and the ability to avoid or reduce animal experimentation.


Osteoblastic Cell Bone Marrow Stromal Cell Mineral Research Osteoblast Lineage Bone Sialoprotein 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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  1. 1.
    Nijweide. P.J., Burger. E.H. and Feyen, J.H. (1986) Cells of bone: proliferation, differentiation, and hormonal regulation. Physiological Reviews 66), 855–886.PubMedGoogle Scholar
  2. 2.
    Rodan, G.A., Heath, J.K., Yoon, K. et al. (1988) Diversity of the osteoblast phenotype, in Cell and Molecular Biology of Vertebrate Hard Tissues, (eds D. Evered and S. Harnett), John Wiley & Sons, Chichester, pp. 78–85.Google Scholar
  3. 3.
    Aubin, J.E., Turksen, K. and Heersche, J.N.M. (1993) Osteoblastic cell lineage, in Cellular and Molecular Biology of Bone, (ed. M. Noda), Academic Press Inc., San Diego, pp. 1–45.Google Scholar
  4. 4.
    Freshney, R.I. (1983) Culture of Animal Cells: a Manual of Basic Technique, Alan R. Liss Inc., New York.Google Scholar
  5. 5.
    Owen, M. (1967) Uptake of [3H] uridine into precursor pools and RNA in osteogenic cells. Journal of Cell Science 2), 39–56.PubMedGoogle Scholar
  6. 6.
    Owen, M. (1970) The origin of bone cells. International Review of Cytology 28), 213–238.PubMedGoogle Scholar
  7. 7.
    Owen, M. and Friedenstein, A.J. (1988) Stromal cells: marrow-derived osteogenic precursors, in Cell and Molecular Biology of Vertebrate Hard Tissues, (eds D. Evered and S. Harnett), John Wiley & Sons, Chichester, pp. 42–53.Google Scholar
  8. 8.
    Wolpert, L. (1988) Stem cells: a problem in assymetry. Journal of Cell Science 10), 1–19.Google Scholar
  9. 9.
    Nakahata, T., Gross, A.J. and Ogawa, M. (1982) A stochastic model of self renewal and commitment to differentiation of the primitive hemopoietic stem cells in culture. Journal of Cellular Physiology 113), 455–458.PubMedCrossRefGoogle Scholar
  10. 10.
    Friedenstein, A.J., Chailalhyan, R.K. and Gerasimov, U.V. (1987) Bone marrow osteogenic stem cells: in vitro cultivation and transplantation in diffusion chambers. Cell and Tissue Kinetics 20), 263–272.PubMedGoogle Scholar
  11. 11.
    Friedenstein, A.J. (1976) Precursor cells of mechanocytes. International Reviews in Cytology 47), 327–359.CrossRefGoogle Scholar
  12. 12.
    Beresford, J.N., Joyner, C.J., Devlin, C. and Triffitt, J.T. (1994) The effects of dexamethasone and 1,25-dihydroxyvitamin D3 on osteogenic differentiation of human marrow stromal cells in vitro. Archives of Oral Biology 39), 941–947.PubMedCrossRefGoogle Scholar
  13. 13.
    Beresford, J.N., Bennett, J.H., Devlin, C. et al. (1992) Evidence for an inverse relation-ship between the differentiation of adipocytic and osteogenic cells in rat marrow stromal cell cultures. Journal of Cell Science 102), 341–351.PubMedGoogle Scholar
  14. 14.
    Maniatopoulos, C., Sodek, J. and Melcher, A.H. (1988) Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats. Cell Tissue Research 254), 317–330.PubMedGoogle Scholar
  15. 15.
    Reznikov, C.A., Brankow, D.W. and Heidelberger, C. (1973) Establishment and characterisation of a cloned line of C3H mouse embryo cells sensitive to postconfluence inhibition of division. Cancer Research 33), 3231–3238.Google Scholar
  16. 16.
    Grigoriadis, A.E., Heersche, J.N.M. and Aubin, J.E. (1988) Differentiation of muscle, fat, cartilage and bone from progenitor cells present in a bone derived clonal cell population: effects of dexamethasone. Journal of Cell Biology 106), 2139–2151.PubMedCrossRefGoogle Scholar
  17. 17.
    Yamaguchi, A. and Kahn, A.J. (1991) Clonal osteogenic cell lines express myogenic and adipocytic developmental potential. Calcified Tissue International 49), 221–225.PubMedCrossRefGoogle Scholar
  18. 18.
    Kellermann, O., Buc-Caron, M.H., Marie, P.J. et al. (1990) An immortalized osteogenic cell line derived from mouse teratocarcinoma is able to mineralize in vivo and in vitro. Journal of Cell Biology 110), 123–132.PubMedCrossRefGoogle Scholar
  19. 19.
    Poliard, A., Lamblin, D., Marie, P.J. et al. (1993) Commitment of the teratocarcinoma-derived mesodermal clone Cl towards terminal osteogenic differentiation. Journal of Cell Science 106), 503–511.PubMedGoogle Scholar
  20. 20.
    Grigoriadis, A.E., Heersche, J.N. and Aubin, J.E. (1990) Continuously growing bipotential and monopotential myogenic, adipogenic, and chondrogenic subclones isolated from the multipotential RCJ 3.1 clonal cell line. Developmental Biology 142), 313–318.PubMedCrossRefGoogle Scholar
  21. 21.
    Konieczny, S.F. and Emerson, C.P. Jr (1984) 5-Azacytidine induction of stable mesodermal stem cell lineages from 10T1/2 cells: evidence for regulatory genes controlling determination. Cell 38), 791–800.PubMedCrossRefGoogle Scholar
  22. 22.
    Turksen, K. and Aubin, J.E. (1991) Positive and negative immunoselection for enrichment of two classes of osteoprogenitor cells. Journal of Cell Biology 114), 373–384.PubMedCrossRefGoogle Scholar
  23. 23.
    Bellows, CG., Aubin, J.E. and Heersche, J.N. (1987) Physiological concentrations of glucocorticoids stimulate formation of bone nodules from isolated rat calvaria cells in vitro. Endocrinology 121), 1985–1992.PubMedGoogle Scholar
  24. 24.
    Hughes, F.J., Collyer, J., Stanfield, M. and Goodman, S.A. (1995) The effects of bone morphogenetic protein-2,-4 and-6 on differentiation of rat osteoblast cells in vitro. Endocrinology 136), 2671–2677.PubMedCrossRefGoogle Scholar
  25. 25.
    McCulloch, C.A.G., Strugurescu, M., Hughes, F. et al. (1991) Osteogenic progenitor cells in rat bone marrow stromal populations exhibit self-renewal in culture. Blood 77), 1906–1911.PubMedGoogle Scholar
  26. 26.
    Kamalia, N., Mcculloch, C.A.G., Tenebaum, H.C. and Limeback, H. (1992) Dexamethasone recruitment of self-renewing osteoprogenitor cells in chick bone marrow stromal cell cultures. Blood 79), 320–326.PubMedGoogle Scholar
  27. 27.
    Bellows, C.G., Heersche, J.N. and Aubin, J.E. (1990) Determination of the capacity for proliferation and differentiation of osteoprogenitor cells in the presence and absence of dexamethasone. Developmental Biology 140), 132–138.PubMedCrossRefGoogle Scholar
  28. 28.
    Aubin, J.E., Gupta, A., Zirngibl, R. and Rossant, J. (1995) Bone sialoprotein knockout mice have bone abnormalities. Bone 17), 558(Abstr. 3a).CrossRefGoogle Scholar
  29. 29.
    Ducy, P., Desbois, C., Boyce, B. et al. (1996) Increased bone formation in osteocalcin-deficient mice. Nature 382), 448–452.PubMedCrossRefGoogle Scholar
  30. 30.
    Young, M.F., Ibaraki, K., Kerr, J.M. and Heegaard, A.-M. (1993) Molecular and cellular biology of the major noncollagenous proteins in bone, in Cellular and Molecular Biology of Bone (ed. M. Noda), Academic Press Inc., San Diego, pp. 191–234.Google Scholar
  31. 31.
    Fisher, L.W. (1985) The nature of the proteoglycans of bone, in The Chemistry and Biology of Mineralised Tissues (ed. W.T. Butler), EBSCO Media, Birmingham, Alabama, pp. 188–196.Google Scholar
  32. 32.
    Noda, M., Yoon, K., Rodan, G.A. and Koppel, D.E. (1987) High lateral mobility of endogenous and transfected alkaline phopshatase: a phosphatidylinositol anchored membrane protein. Journal of Cell Biology 105), 1671–1677.PubMedCrossRefGoogle Scholar
  33. 33.
    Modrowski, D. and Marie, P.J. (1993) Cells isolated from the endosteal bone surface of adult rats express differentiated osteoblastic characteristics in vitro. Cell and Tissue Research 271), 499–505.PubMedGoogle Scholar
  34. 34.
    Beresford, J.N., Gallagher, J.A., Poser, J.W. and Russell, R.G. (1984) Production of osteocalcin by human bone cells in vitro. Effects of 1,25(OH)2D3, 24,25(OH)2D3, parathyroid hormone, and glucocorticoids. Metabolic Bone Disease and Related Research 5), 229–234.CrossRefGoogle Scholar
  35. 35.
    Murray, E., Provvedini, D., Curran, D. et al. (1987) Characterization of a human osteoblastic osteosarcoma cell line (SaOS-2) with high bone alkaline phosphatase activity. Journal of Bone and Mineral Research 2), 231–238.PubMedGoogle Scholar
  36. 36.
    Partridge, N.C., Alcorn, D., Michelangeli, V.P. et al. (1983) Morphological and biochemical characterization of four clonal osteogenic sarcoma cell lines of rat origin. Cancer Research 43), 4308–314.PubMedGoogle Scholar
  37. 37.
    Rao, L.G., Ng, B., Brunette, D.M. and Heersche, J.N.M. (1977) Parathyroid hormone and prostaglandin E1 response in a selected population of bone cells after repeated subculture and storage at-80°C. Endocrinology 100), 1233–1241.PubMedGoogle Scholar
  38. 38.
    Robey, P.G. and Termine, J.D. (1985) Human bone cells in vitro. Calcified Tissue International 37), 453–460.PubMedCrossRefGoogle Scholar
  39. 39.
    Rodan, S.B., Imai, Y., Thiede, M.A. et al. (1987) Characterization of a human osteosarcoma cell line (Saos-2) with osteoblastic properties. Cancer Research 47), 4961–4966.PubMedGoogle Scholar
  40. 40.
    Akatsu, T., Takahashi, N., Udagawa, N. et al. (1991) Role of prostaglandins in interleukin-1-induced bone resorption in mice in vitro. Journal of Bone and Mineral Research 6), 183–189.PubMedGoogle Scholar
  41. 41.
    Ellies, L.G. and Aubin, J.E. (1990) Temporal sequence of interleukin 1 alpha-mediated stimulation and inhibition of bone formation by isolated fetal rat calvaria cells in vitro. Cytokine 2), 430–437.PubMedCrossRefGoogle Scholar
  42. 42.
    Evans, D.B., Thavarajah, M. and Kanis, J.A. (1990) Involvement of prostaglandin E2 in the inhibition of osteocalcin synthesis by human osteoblast-like cells in response to cytokines and systemic hormones. Biochemical and Biophysical Research Communications 167), 194–202.PubMedCrossRefGoogle Scholar
  43. 43.
    Saito, S., Ngan, P., Rosol, T. et al. (1991) Involvement of PGE synthesis in the effect of intermittent pressure and interleukin-1 beta on bone resorption. Journal of Dental Research 70), 27–33.PubMedGoogle Scholar
  44. 44.
    Schwartz, Z., Dennis, R., Bonewald, L. et al. (1992) Differential regulation of prostaglandin E2 synthesis and phospholipase A2 activity by 1,25-(OH)2D3 in three osteoblast-like cell lines (MC-3T3-E1, ROS 17/2.8, and MG-63). Bone 13), 51–58.PubMedCrossRefGoogle Scholar
  45. 45.
    Fried, A., Benayahu, D. and Wientroub, S. (1993) Marrow stroma-derived osteogenic clonal cell lines: putative stages in osteoblastic differentiation. Journal of Cellular Physiology 155), 472–82.PubMedCrossRefGoogle Scholar
  46. 46.
    Kasugai, S., Oida, S., Iimura, T. et al. (1995) Expression of prostaglandin E receptor subtypes in bone: expression of EP2 in bone development. Bone 17), 1–4.PubMedCrossRefGoogle Scholar
  47. 47.
    Scutt, A. and Bertram, P. (1995) Bone marrow cells are targets for the anabolic actions of prostaglandin E2 on bone: induction of a transition from nonadherent to adherent osteoblast precursors. Journal of Bone and Mineral Research 10), 474–487.PubMedGoogle Scholar
  48. 48.
    Evans, D.B., Bunning, R.A., Van Damme, J. and Russell, R.G. (1989) Natural human IL-1 beta exhibits regulatory actions on human bone-derived cells in vitro. Biochemical and Biophysical Research Communications 159), 1242–1248.PubMedCrossRefGoogle Scholar
  49. 49.
    Evans, D.B., Bunning, R.A. and Russell, R.G. (1990) The effects of recombinant human interleukin-1 beta on cellular proliferation and the production of prostaglandin E2, plasminogen activator, osteocalcin and alkaline phosphatase by osteoblast-like cells derived from human bone. Biochemical and Biophysical Research Communications 166), 208–216.PubMedCrossRefGoogle Scholar
  50. 50.
    Hanazawa, S., Ohmori, Y., Amano, S. et al. (1986) Human purified interleukin-1 inhibits DNA synthesis and cell growth of osteoblastic cell line (MC3T3-E1), but enhances alkaline phosphatase activity in the cells. FEBS Letters 203), 279–284.PubMedCrossRefGoogle Scholar
  51. 51.
    Ikeda, E., Kusaka, M., Hakeda, Y. et al. (1988) Effect of interleukin 1 beta on osteoblastic clone MC3T3-E1 cells. Calcified Tissue International 43), 162–166.PubMedCrossRefGoogle Scholar
  52. 52.
    Jin, C.H., Miyaura, C., Ishimi, Y. et al. (1990) Interleukin 1 regulates the expression of osteopontin mRNA by osteoblasts. Molecular and Cellular Endocrinology 74), 221–228.PubMedCrossRefGoogle Scholar
  53. 53.
    Gowen, M., Wood, D.D. and Russell, R.G. (1985) Stimulation of the proliferation of human bone cells in vitro by human monocyte products with interleukin-1 activity. Journal of Clinical Investigation 75), 1223–1229.PubMedGoogle Scholar
  54. 54.
    Kuroki, T., Shingu, M., Koshihara, Y. and Nobunaga, M. (1994) Effects of cytokines on alkaline phosphatase and osteocalcin production, calcification and calcium release by human osteoblastic cells. British Journal of Rheumatology 33), 224–230.PubMedCrossRefGoogle Scholar
  55. 55.
    Rickard, D.J., Gowen, M. and MacDonald, B.R. (1993) Proliferative responses to estradiol, IL-1 alpha and TGF beta by cells expressing alkaline phosphatase in human osteoblast-like cell cultures. Calcified Tissue International 52), 227–233.PubMedCrossRefGoogle Scholar
  56. 56.
    Stashenko, P., Dewhirst, F.E., Rooney, M.L. et al. (1987) Interleukin-1 beta is a potent inhibitor of bone formation in vitro. Journal of Bone and Mineral Research 2), 559–565.PubMedGoogle Scholar
  57. 57.
    Antosz, M.E., Bellows, C.G. and Aubin, J.E. (1989) Effects of transforming growth factor ß and epidermal growth factor on cell proliferation and the formation of bone nodules in isolated fetal rat calvaria cells. Journal of Cellular Physiology 140), 386–395.PubMedCrossRefGoogle Scholar
  58. 58.
    Clover, J. and Gowen, M. (1994) Are MG-63 and HOS TE85 human osteosarcoma cell lines representative models of the osteoblastic phenotype? Bone 15), 585–591.PubMedCrossRefGoogle Scholar
  59. 59.
    Grzesik, W.J. and Robey, P.G. (1994) Bone matrix RGD glycoproteins: immunolocal-ization and interaction with human primary osteoblastic bone cells in vitro. Journal of Bone and Mineral Research 9), 487–496.PubMedGoogle Scholar
  60. 60.
    Hughes, D.E., Salter, D.M., Dedhar, S. and Simpson, R. (1993) Integrin expression in human bone. Journal of Bone and Mineral Research 8), 527–533.PubMedGoogle Scholar
  61. 61.
    Hughes, D.E., Salter, D.M. and Simpson, R. (1994) CD44 expression in human bone: a novel marker of osteocytic differentiation. Journal of Bone and Mineral Research 9), 39–4.PubMedGoogle Scholar
  62. 62.
    Horowitz, M.C., Fields, A., DeMeo, D. et al. (1994) Expression and regulation of Ly-6 differentiation antigens by murine osteoblasts. Endocrinology 135), 1032–1043.PubMedCrossRefGoogle Scholar
  63. 63.
    Jamal, H.H. and Aubin, J.E. (1996) CD44 expression in fetal rat bone: in vivo and in vitro analysis. Experimental Cell Research 223), 467–477.PubMedCrossRefGoogle Scholar
  64. 64.
    Nakamura, H., Kenmotsu, S., Sakai, H. and Ozawa, H. (1995) Localization of CD44, the hyaluronate receptor, on the plasma membrane of osteocytes and osteoclasts in rat tibiae. Cell and Tissue Research 280), 225–233.PubMedGoogle Scholar
  65. 65.
    Pavasant P., Shizari, T.M. and Underhill, C.B. (1994) Distribution of hyaluronan in the epiphysial growth plate: turnover by CD44-expressing osteoprogenitor cells. Journal of Cell Science 107), 2669–2677.PubMedGoogle Scholar
  66. 66.
    Van Vlasselaer, P., Falla, N., Snoeck, H. and Mathieu, E. (1994) Characterization and purification of osteogenic cells from murine bone marrow by two-color cell sorting using anti-Sca-1 monoclonal antibody and wheat germ agglutinin. Blood 84), 753–63.Google Scholar
  67. 67.
    Simmons, P.J. and Torok-Storb, B. (1991) Identification of stromal cell precursors in human bone marrow by a novel monoclonal antibody, STRO-1. Blood 78), 55–62.PubMedGoogle Scholar
  68. 68.
    Gronthos, S., Graves, S.E., Ohta, S. and Simmons, P.J. (1994) The STRO-1+ fraction of adult human bone marrow contains the osteogenic precursors. Blood 84), 4164–4173.PubMedGoogle Scholar
  69. 69.
    Nakamura, T., Gross, M., Yamamuro, T. and Liao, S.-K. (1987) Identification of a human osteosarcoma-associated glycoprotein with monoclonal antibodies: relationship with alkaline phosphatase. Biochemistry and Cell Biology 65), 1091–1097.PubMedCrossRefGoogle Scholar
  70. 70.
    Bruder, S.P. and Caplan, A.I. (1990) Osteogenic cell lineage analysis is facilitated by organ cultures of embryonic chick periosteum. Developmental Biology 141), 319–329.PubMedCrossRefGoogle Scholar
  71. 71.
    Turksen, K., Bhargava, U., Moe, H.K. and Aubin, J.E. (1992) Isolation of monoclonal antibodies recognising rat bone-associated molecules in vivo and in vitro. Journal of Histochemistry and Cytochemistry 40), 1339–1352.PubMedGoogle Scholar
  72. 72.
    Yamaguchi, A. and Kahn, A.J. (1993) Monoclonal antibodies that recognize antigens in human osteosarcoma cells and normal fetal osteoblasts. Bone and Mineral 22), 165–176.PubMedGoogle Scholar
  73. 73.
    Aubin, J.E. and Turksen, K. (1996) Monoclonal antibodies as tools for studying the osteoblast lineage. Microscopy Research and Technique 33), 128–140.PubMedCrossRefGoogle Scholar
  74. 74.
    Nijweide, P.J. and Mulder, R.J. (1986) Identification of osteocytes in osteoblast-like cell cultures using a monoclonal antibody specifically directed against osteocytes. Histochemistry 84), 342–347.PubMedCrossRefGoogle Scholar
  75. 75.
    Bruder, S.P. and Caplan, A.I. (1989) First bone formation and the dissection of an osteogenic lineage in the embryonic chick tibia is revealed by monoclonal antibodies against osteoblasts. Bone 10), 359–35.PubMedCrossRefGoogle Scholar
  76. 76.
    van der Plas, A. and Nijweide, P.J. (1992) Isolation and purification of osteocytes. Journal of Bone and Mineral Research 7), 389–396.PubMedGoogle Scholar
  77. 77.
    van der Plas, A., Aarden, E.M., Feijen, J.H. et al. (1994) Characteristics and properties of osteocytes in culture. Journal of Bone and Mineral Research 9), 1697–1704.PubMedGoogle Scholar
  78. 78.
    Wetterwald, A. and Fleisch, H. (1990) A monoclonal antibody recognises a cell membrane antigen present in a subpopulation of rat osteoblasts and in osteocytes. Calcified Tissue International 46(Suppl. 2), A13 (Abstr. 52).Google Scholar
  79. 79.
    Nose, K., Saito, H. and Kuroli, T. (1990) Isolation of a gene sequence induced later by tumor-promoting 12-O-tetradecanolyphorbol-13-acetate im mouse osteoblastic cells (MC3T3-E1) and expressed constitutively in ras transformed cells. Cell Growth and Differentiation 1), 511–518.PubMedGoogle Scholar
  80. 80.
    Wetterwald, A., Hofstetter, W., Cecchini, M.G. et al. (1996) Characterisation and cloning of the E11 antigen, a marker expressed by rat osteoblasts and osteocytes. Bone 18), 125–132.PubMedCrossRefGoogle Scholar
  81. 81.
    Sprague, L., Wetterwald, A., Heinzman, U. and Atkinson, MJ. (1996) Phenotypic changes following over-expression of sense of antisense E11 cDNA in ROS 17/2.8 cells. Journal of Bone and Mineral Research 11), S132.Google Scholar
  82. 82.
    Haynesworth, S.E., Baber, M.A. and Caplan, A.I. (1992) Cell surface antigens on human marrow-derived mesenchymal cells are detected by monoclonal antibodies. Bone 13), 69–80.PubMedCrossRefGoogle Scholar
  83. 83.
    Bellows, C.G., Aubin, J.E. and Heersche, J.N. (1991) Initiation and progression of mineralization of bone nodules formed in vitro: the role of alkaline phosphatase and organic phosphate. Bone and Mineral 14), 27–40.PubMedCrossRefGoogle Scholar
  84. 84.
    Bellows, C.G., Heersche, J.N. and Aubin, J.E. (1992) Inorganic phosphate added exogenously or released from beta-glycerophosphate initiates mineralization of osteoid nodules in vitro. Bone and Mineral 17), 15–29.PubMedCrossRefGoogle Scholar
  85. 85.
    Marsh, M.E., Munne, A.M., Vogel, J.J. et al. (1995) Mineralization of bone-like extracellular matrix in the absence of functional osteoblasts. Journal of Bone and Mineral Research 10), 1635–1643.PubMedGoogle Scholar
  86. 86.
    Beertsen, W. and Vandenbos, T. (1992) Alkaline phosphatase induces the mineralization of sheets of collagen implanted subcutaneously in the rat. Journal of Clinical Investigation 89), 1974–1980.PubMedGoogle Scholar
  87. 87.
    Khouja, H.I., Bevington, A., Kemp, G.J. and Russell, R.G.G. (1990) Calcium and orthophosphate deposits in vitro do not imply osteoblast mediated mineralization: mineralization by betaglycerophosphate in the absence of osteoblasts. Bone 11), 385–391.PubMedCrossRefGoogle Scholar
  88. 88.
    Bellows, CG., Aubin, J.E., Heersche, J.N.M. and Antosz, M.E. (1986) Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations. Calcified Tissue International 38,, 143–154.PubMedCrossRefGoogle Scholar
  89. 89.
    Bhargava, U., Bar-Lev, M., Bellows, C.G. and Aubin, J.E. (1988) Ultrastructural analysis of bone nodules formed in vitro by isolated fetal rat calvaria cells. Bone 9), 155–163.PubMedCrossRefGoogle Scholar
  90. 90.
    Ecarot-Charrier, B., Glorieux, F.H., van der Rest, M. and Pereira, G. (1983) Osteoblasts isolated from mouse calvaria initiate matrix mineralization in culture. Journal of Cell Biology 96), 639–643.PubMedCrossRefGoogle Scholar
  91. 91.
    Nefussi, J.R., Boy-Lefevre, M.L., Boulekbache, H. and Forest, N. (1985) Mineralization in vitro of matrix formed by osteoblasts isolated by collagenase digestion. Differentiation 29), 160–168.PubMedCrossRefGoogle Scholar
  92. 92.
    Long, M.W., Robinson, J.A., Ashcraft, E.A. and Mann, K.G. (1995) Regulation of human bone marrow-derived osteoprogenitor cells by osteogenic growth factors. Journal of Clinical Investigation 95), 881–887.PubMedGoogle Scholar
  93. 93.
    Lee, K.L., Aubin, J.E. and Heersche, J.N. (1992) Beta-glycerophosphate-induced mineralization of osteoid does not alter expression of extracellular matrix components in fetal rat calvarial cell cultures. Journal of Bone and Mineral Research 7), 1211–1219.PubMedGoogle Scholar
  94. 94.
    Rao, L.G., Murray, T.M. and Heersche, J.N. (1983) Immunohistochemical demonstration of parathyroid hormone binding to specific cell types in fixed rat bone tissue. Endocrinology 113), 805–810.PubMedGoogle Scholar
  95. 95.
    Silve, CM., Hradek, G.T., Jones, A.L. and Arnaud, C.D. (1982) Parathyroid hormone receptor in intact embryonic chicken bone: characterization and cellular localization. Journal of Cell Biology 94), 379–386.PubMedCrossRefGoogle Scholar
  96. 96.
    Rouleau, M.F., Mitchell, J. and Goltzman, D. (1988) In vivo distribution of parathyroid hormone receptors in bone: evidence that a predominant osseous target cell is not the mature osteoblast. Endocrinology 123), 187–191.PubMedGoogle Scholar
  97. 97.
    Rouleau, M.F., Mitchell, J. and Goltzman, D. (1990) Characterization of the major parathyroid hormone target cell in the endosteal metaphysis of rat long bones. Journal of Bone and Mineral Research 5), 1043–1–53.PubMedGoogle Scholar
  98. 98.
    Heath, J.K., Rodan, S.B., Yoon, K.G. and Rodan, G.A. (1989) Rat calvaria cell lines immortalised by SV40 large T-antigen-constitutive and retinoic acid-inducible expression of osteoblastic features. Endocrinology 124), 3060–3068.PubMedGoogle Scholar
  99. 99.
    Shalhoub, V., Conlon, D., Tassinari, M. et al. (1992) Glucocorticoids promote development of the osteoblast phenotype by selectively modulating expression of cell growth and differentiation associated genes. Journal of Cellular Biochemistry 50), 425–140.PubMedCrossRefGoogle Scholar
  100. 100.
    Owen, T.A., Aronow, M., Shalhoub, V. et al. (1990) Progressive development of the rat osteoblast phenotype in vitro: reciprocal relationships in expression of genes associated with osteoblast proliferation and differentiation during formation of the bone extracellular matrix. Journal of Cellular Physiology 143), 420–430.PubMedCrossRefGoogle Scholar
  101. 101.
    Owen, T.A., Aronow, M.S., Barone, L.M. et al. (1991) Pleiotropic effects of vitamin D on osteoblast gene expression are related to the proliferative and differentiated state of the bone cell phenotype: dependency upon basal levels of gene expression, duration of exposure, and bone matrix competency in normal rat osteoblast cultures. Endocrinology 128), 1496–1504.PubMedGoogle Scholar
  102. 102.
    Aronow, M.A., Gerstenfeld, L.C., Owen, T.A. et al. (1990) Factors that promote progressive development of the osteoblast phenotype in cultured fetal rat calvaria cells. Journal of Cellular Physiology 143), 213–221.PubMedCrossRefGoogle Scholar
  103. 103.
    Liu, F., Malaval, L., Gupta, A.K. and Aubin, J.E. (1994) Simultaneous detection of multiple bone-related mRNAs and protein expression during osteoblast differentiation: polymerase chain reaction and immunocytochemical studies at the single cell level. Developmental Biology 166), 220–234.PubMedCrossRefGoogle Scholar
  104. 104.
    Breen. E.C., Ignotz, R.A., McCabe, L. et al. (1994) TGF beta alters growth and differentiation related gene expression in proliferating osteoblasts in vitro, preventing development of the mature bone phenotype. Journal of Cellular Physiology 160), 323–335.PubMedCrossRefGoogle Scholar
  105. 105.
    Stein, G.S. and Lian, J.B. (1993) Molecular mechanisms mediating proliferation/differentiation interrelationships during progressive development of the osteoblast phenotype. [Review]. Endocrine Reviews 14), 424–442.PubMedCrossRefGoogle Scholar
  106. 106.
    Smith, E., Frenkel, B., Schlegel, R. et al. (1995) Expression of cell cycle regulatory factors in differentiating osteoblasts: postproliferative up-regulation of cyclins B and E. Cancer Research 55), 5019–5024.PubMedGoogle Scholar
  107. 107.
    Tsuji, T., Hughes, F.J., McCulloch, C.A.G. and Melcher, A.H. (1990) The effect of donor age on the osteogenic cells of rat bone marrow. Mechanisms of Ageing and Development 51), 121–132.PubMedCrossRefGoogle Scholar
  108. 108.
    Lennon, D.P., Haynesworth, S.E., Young, R.G. et al. (1995) A chemically denned medium supports in vitro proliferation and maintains the osteochondral potential of rat marrow-derived mesenchymal stem cells. Experimental Cell Research 219), 211–222.PubMedCrossRefGoogle Scholar
  109. 109.
    Denis, I., Pointillart, A. and Lieberherr, M. (1994) Cell stage-dependent effects of ascorbic acid on cultured porcine bone cells. Bone & Mineral 25), 149–161.Google Scholar
  110. 110.
    Franceschi, R.T., Iyer, B.S. and Cui, Y. (1994) Effects of ascorbic acid on collagen matrix formation and osteoblast differentiation in murine MC3T3-E1 cells. Journal of Bone and Mineral Research 9), 843–854.PubMedGoogle Scholar
  111. 111.
    Harada, S., Matsumoto, T. and Ogata, E. (1991) Role of ascorbic acid in the regulation of proliferation in osteoblast-like MC3T3-E1 cells. Journal of Bone and Mineral Research 6), 903–908.PubMedGoogle Scholar
  112. 112.
    Malaval, L., Modrowski, D., Gupta, A.K. and Aubin, J.E. (1994) Cellular expression of bone-related proteins during in vitro osteogenesis in rat bone marrow stromal cell cultures. Journal of Cellular Physiology 158), 555–572.PubMedCrossRefGoogle Scholar
  113. 113.
    Quarles, L.D., Yohay, D.A., Lever, L.W. et al. (1992) Distinct proliferative and differentiated stages of murine MC3T3-E1 cells in culture: an in vitro model of osteoblast development. Journal of Bone and Mineral Research 7), 683–692.PubMedGoogle Scholar
  114. 114.
    Torii, Y., Hitomi, K. and Tsukagoshi, N. (1994) L-Ascorbic acid 2-phosphate promotes osteoblastic differentiation of MC3T3-E1 mediated by accumulation of type I collagen. Journal of Nutritional Science and Vitaminology 40), 229–238.PubMedGoogle Scholar
  115. 115.
    Dixon, S.J. and Wilson, J.X. (1992) Adaptive regulation of ascorbate transport in osteoblastic cells. Journal of Bone and Mineral Research 7), 675–681.PubMedGoogle Scholar
  116. 116.
    Franceschi, R.T., Wilson, J.X. and Dixon, S.J. (1995) Requirement for Na(+)-dependent ascorbic acid transport in osteoblast function. American Journal of Physiology 268), C1430–C1439.PubMedGoogle Scholar
  117. 117.
    Boskey, A.L., Stiner, D., Doty, S.B. and Binderman, I. (1991) Requirement of vitamin C for cartilage calcification in a differentiating chick limb-bud mesenchymal cell culture. Bone 12), 277–282.PubMedCrossRefGoogle Scholar
  118. 118.
    Beresford, J.N., Graves, S.E. and Smoothy, C.A. (1993) Formation of mineralized nodules by bone derived cells in vitro: a model of bone formation? American Journal of Medical Genetics 45), 163–178.PubMedCrossRefGoogle Scholar
  119. 119.
    McCulloch, C.A.G. and Tenenbaum, H.C. (1986) Dexamethasone induces proliferation and terminal differentiation of osteogenic cells in tissue culture. Anatomical Record 215), 397.PubMedCrossRefGoogle Scholar
  120. 120.
    Ogata, Y., Yamauchi, M., Kim, R.H. et al. (1995) Glucocorticoid regulation of bone sialoprotein (BSP) gene expression. Identification of a glucocorticoid response element in the bone sialoprotein gene promoter. European Journal of Biochemistry 230), 183–192.PubMedCrossRefGoogle Scholar
  121. 121.
    Majeska, R.J., Nair, B.C. and Rodan, G.A. (1985) Glucocorticoid regulation of alkaline phosphatase in the osteoblastic osteosarcoma cell line ROS 17/2.8. Endocrinology 116), 170–179.PubMedGoogle Scholar
  122. 122.
    Takuwa, Y., Ohse, C., Wang, E.A. et al. (1991) Bone morphogenetic protein-2 stimulates alkaline phosphatase activity and collagen synthesis in cultured osteoblastic cells, MC3T3-E1. Biochemical and Biophysical Research Communications 174), 96–101.PubMedCrossRefGoogle Scholar
  123. 123.
    Falla, N., Van, V., Bierkens, J. et al. (1993) Characterization of a 5-fluorouracil-enriched osteoprogenitor population of the murine bone marrow. Blood 82), 3580–3591.PubMedGoogle Scholar
  124. 124.
    Herbertson, A. and Aubin, J.E. (1995) Dexamethasone alters the subpopulation makeup of rat bone marrow stromal cell cultures. Journal of Bone and Mineral Research 10), 285–294.PubMedGoogle Scholar
  125. 125.
    Benayahu, D., Kletter, Y., Zipori, D. and Wientroub, S. (1989) Bone marrow-derived stromal cell line expressing osteoblastic phenotype in vitro and osteogenic capacity in vivo. Journal of Cellular Physiology 140), 1–7.PubMedCrossRefGoogle Scholar
  126. 126.
    Gundle, R. and Beresford, J.N. (1995) The isolation and culture of cells from expiants of human trabecular bone. Calcified Tissue International 56), S8–S10.PubMedGoogle Scholar
  127. 127.
    Whitson, S.W., Harrison, W., Dunlap, M.K. et al. (1984) Fetal bovine bone cells synthesize bone-specific matrix proteins. Journal of Cell Biology 99), 607–614.PubMedCrossRefGoogle Scholar
  128. 128.
    Wong, G.L. and Cohn, D.V. (1975) Target cells in bone for parathormone and calcitonin are different: enrichment for each cell type by sequential digestion of mouse calvaria and selective adhesion to polymeric surfaces. Proceedings of the National Academy of Sciences USA 72), 3167–3171.CrossRefGoogle Scholar
  129. 129.
    Nijweide, P.J., van Iperen-van Gent, A.S., Kawilarang-de Haas E.W. et al. (1982) Bone formation and calcification by isolated osteoblastlike cells. Journal of Cell Biology 93), 318–323.PubMedCrossRefGoogle Scholar
  130. 130.
    Bellows, C.G. and Aubin, J.E. (1989) Determination of numbers of osteoprogenitor cells present in isolated fetal rat calvaria cells in vitro. Developmental Biology 133), 8–13.Google Scholar
  131. 131.
    Bellows, C.G., Ishida, H., Aubin, J.E. and Heersche, J.N. (1990) Parathyroid hormone reversibly suppresses the differentiation of osteoprogenitor cells into functional osteoblasts. Endocrinology 127), 3111–3116.PubMedCrossRefGoogle Scholar
  132. 132.
    Harris, S.E., Bonewald, L.F., Harris, M.A. et al. (1994) Effects of transforming growth factor beta on bone nodule formation and expression of bone morphogenetic protein 2, osteocalcin, osteopontin, alkaline phosphatase, and type I collagen mRNA in long-term cultures of fetal rat calvarial osteoblasts. Journal of Bone and Mineral Research 9), 855–863.PubMedGoogle Scholar
  133. 133.
    Hughes, F.J. and Howells, G.L. (1993) Interleukin-11 inhibits bone formation in vitro. Calcified Tissue International 53), 362–364.PubMedCrossRefGoogle Scholar
  134. 134.
    Hughes, F.J. and Howells, G.L. (1993) Interleukin-6 inhibits bone formation in vitro. Bone and Mineral 21), 21–28.PubMedGoogle Scholar
  135. 135.
    Leboy, P.S., Beresford, J.N., Devlin, C. and Owen, M.E. (1991) Dexamethasone induction of osteoblast mRNAs in rat marrow stromal cell cultures. Journal of Cellular Physiology 146), 370–378.PubMedCrossRefGoogle Scholar
  136. 136.
    Hughes, F.J. and McCulloch, C.A.G. (1991) Stimulation of the differentiation of osteogenic rat bone marrow stromal cells by osteoblast cultures. Laboratory Investigation 64), 617–622.PubMedGoogle Scholar
  137. 137.
    Taylor, S.M. and Jones, P.A. (1979) Multiple new phenotypes induced in 10T1/2 and 3T3 cells treated with 5-azacytidine. Cell 17), 771–779.PubMedCrossRefGoogle Scholar
  138. 138.
    Taylor, S.M. and Jones, P.A. (1982) Changes in phenotypic expression in embryonic and adult cells treated with 5-azacytidine. Journal of Cellular Physiology 111), 187–194.PubMedCrossRefGoogle Scholar
  139. 139.
    Wang, E.A., Israel, D.I., Kelly, S. and Luxenberg, D.P. (1993) Bone morphogenetic protein-2 causes commitment and differentiation in C3H10T1/2 and 3T3 cells. Growth Factors 9), 57–71.PubMedGoogle Scholar
  140. 140.
    Ahrens, M., Ankenbauer, T., Schroder, D. et al. (1993) Expression of human bone morphogenetic proteins-2 or-4 in murine mesenchymal progenitor C3H10T1/2 cells induces differentiation into distinct mesenchymal cell lineages. DNA and Cell Biology 12), 871–880.PubMedGoogle Scholar
  141. 141.
    Katagiri, T., Yamaguchi, A., Ikeda, T. et al. (1990) The non-osteogenic mouse pluripotent cell line, C3H10T1/2, is induced to differentiate into osteoblastic cells by recombinant human bone morphogenetic protein-2. Biochemical and Biophysical Research Communications 172), 295–299.PubMedCrossRefGoogle Scholar
  142. 142.
    Aubin, J.E., Heersche, J.N.M., Merrilees, M.J. and Sodek, J. (1982) Isolation of bone cell clones with differences in growth, hormone responses, and extracellular matrix produc-tion. Journal of Cell Biology 92), 452–461.PubMedCrossRefGoogle Scholar
  143. 143.
    Gitelman, S.E., Kirk, M., Ye, J.Q. et al. (1995) Vgr-1/BMP-6 induces osteoblastic differentiation of pluripotential mesenchymal cells. Cell Growth & Differentiation 6), 827–836.Google Scholar
  144. 144.
    Yamaguchi, A., Katagiri, T., Ikeda, T. et al. (1991) Recombinant human bone morphogenetic protein-2 stimulates osteoblastic maturation and inhibits myogenic differentiation in vitro. Journal of Cell Biology 113), 681–687.PubMedCrossRefGoogle Scholar
  145. 145.
    Poliard, A., Nifuji, A., Lamblin, D. et al. (1995) Controlled conversion of an immortalized mesodermal progenitor cell towards osteogenic, chondrogenic, or adipogenic pathways. Journal of Cell Biology 130), 1461–1472.PubMedCrossRefGoogle Scholar
  146. 146.
    Chentoufi, J., Hott, M., Lamblin, D. et al. (1993) Kinetics of in vitro mineralization by an osteogenic clonal cell line (C1) derived from mouse teratocarcinoma. Differentiation 53), 181–189.PubMedCrossRefGoogle Scholar
  147. 147.
    Asahina, I., Sampath, T.K. and Hauschka, P.V. (1996) Human osteogenic protein-1 induces chondroblastic, osteoblastic, and/or adipocytic differentiation of clonal murine target cells. Experimental Cell Research 222), 38–47.PubMedCrossRefGoogle Scholar
  148. 148.
    Brennan, T.J., Edmondson, D.G., Li, L. and Olson, E.N. (1991) Transforming growth factor beta represses the actions of myogenin through a mechanism independent of DNA binding. Proceedings of the National Academy of Sciences USA 88), 3822–3826.CrossRefGoogle Scholar
  149. 149.
    Kodama, H., Amagai, Y., Sudo, H. et al. (1982) Establishment of a clonal osteogenic cell line from newborn mouse calvaria. Japanese Journal of Oral Biology 23), 899–904.Google Scholar
  150. 150.
    Sudo, H., Kodama, H., Amagai, Y. et al. (1983) In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria. Journal of Cell Biology 96), 191–198.PubMedCrossRefGoogle Scholar
  151. 151.
    Benayahu, D., Fried, A., Shamay, A. et al. (1994) Differential effects of retinoic acid and growth factors on osteoblastic markers and CD10/NEP activity in stromal-derived osteoblasts. Journal of Cellular Biochemistry 56), 62–73.PubMedCrossRefGoogle Scholar
  152. 152.
    Benayahu, D., Kompier, R., Shamay, A. et al. (1994) Mineralization of marrow-stromal osteoblasts MBA-15 on three-dimensional carriers. Calcified Tissue International 55), 120–127.PubMedCrossRefGoogle Scholar
  153. 153.
    Benayahu, D., Fried, A. and Wientroub, S. (1995) PTH and 1,25(OH)2 vitamin D priming to growth factors differentially regulates the osteoblastic markers in MBA-15 clonal subpopulations. Biochemical and Biophysical Research Communications 210), 197–204.PubMedCrossRefGoogle Scholar
  154. 154.
    Sodek, J., Kim, R.H., Ogata, Y. et al. (1995) Regulation of bone sialoprotein gene transcription by steroid hormones. Connective Tissue Research 32), 209–217.PubMedGoogle Scholar
  155. 155.
    Yoon, K.G., Rutledge, S.J., Buenaga, R.F. and Rodan, G.A. (1988) Characterization of the rat osteocalcin gene: stimulation of promoter activity by 1,25-dihydroxyvitamin D3. Biochemistry 27), 8521–8526.PubMedCrossRefGoogle Scholar
  156. 156.
    Franceschi, R.T. and Young, J. (1990) Regulation of alkaline phosphatase by 1,25-dihy-droxyvitamin D3 and ascorbic acid in bone-derived cells. Journal of Bone and Mineral Research 5), 1157–1167.PubMedGoogle Scholar
  157. 157.
    Zhou, H., Hammonds, R. Jr, Findlay, D.M. et al. (1993) Differential effects of transforming growth factor-beta 1 and bone morphogenetic protein 4 on gene expression and differentiated function of preosteoblasts. Journal of Cellular Physiology 155), 112–119.PubMedCrossRefGoogle Scholar
  158. 158.
    Ng, K.W., Gummer, P.R., Michelangeli, V.P. et al. (1988) Regulation of alkaline phosphatase expression in a neonatal rat clonal calvarial cell strain by retinoic acid. Journal of Bone and Mineral Research 3), 53–61.PubMedGoogle Scholar
  159. 159.
    Forrest, S.M., Ng, K.W., Findlay, D.M. et al. (1985) Characterization of an osteoblast-like clonal cell line which responds to both parathyroid hormone and calcitonin. Calcified Tissue International 37), 51–56.PubMedCrossRefGoogle Scholar
  160. 160.
    Ferrier, J., Ward-Kesthely, A., Heersche, J.N.M. and Aubin, J.E. (1989) Membrane potential changes, cAMP stimulation and contraction in osteoblast-like UMR 106 cells in response to calcitonin and parathyroid hormone. Bone and Mineral 4), 133–145.Google Scholar
  161. 161.
    Randall, J.C., Morris, D.C., Zeiger, S. et al. (1989) Presence and activity of alkaline phosphatase in two human osteosarcoma cell lines. Journal of Histochemistry and Cytochemistry 37), 1069–1074.PubMedGoogle Scholar
  162. 162.
    McQuillan, D.J., Richardson, M.D. and Bateman, J.F. (1995) Matrix deposition by a calcifying human osteogenic sarcoma cell line (SaOS-2). Bone 16), 415–426.PubMedGoogle Scholar
  163. 163.
    Farley, J.R., Kyeyune-Nyombi, E., Tarbaux, N.M. et al. (1989) Alkaline phosphatase activity from human osteosarcoma cell line SaOS-2: an isoenzyme standard for quantifying skeletal alkaline phosphatase activity in serum. Clinical Chemistry 35), 223–229.PubMedGoogle Scholar
  164. 164.
    Jaaskelainen, T., Pirskanen, A., Ryhanen, S. et al. (1994) Functional interference between AP-1 and the vitamin D receptor on osteocalcin gene expression in human osteosarcoma cells. European Journal of Biochemistry 224), 11–20.PubMedCrossRefGoogle Scholar
  165. 165.
    Billiau, A., Edy, V.G., Heremans, H. et al. (1977) Human interferon: mass production in a newly established cell line, MG-63. Antimicrobial Agents and Chemotherapy 12), 11–15.PubMedGoogle Scholar
  166. 166.
    Franceschi, R.T., James, W.M. and Zerlauth, G. (1985) 1 alpha, 25-dihydroxyvitamin D3 specific regulation of growth, morphology, and fibronectin in a human osteosarcoma cell line. Journal of Cellular Physiology 123), 401–409.PubMedCrossRefGoogle Scholar
  167. 167.
    Dedhar, S., Mitchell, M.D. and Pierschbacher, M.D. (1989) The osteoblast-like differentiated phenotype of a variant of MG-63 osteosarcoma cell line correlated with altered adhesive properties. Connective Tissue Research 20), 49–61.PubMedGoogle Scholar
  168. 168.
    Lajeunesse, D., Kiebzak, G.M., Frondoza, C. and Sacktor, B. (1991) Regulation of osteocalcin secretion by human primary bone cells and by the human osteosarcoma cell line MG-63. Bone and Mineral 14), 237–250.PubMedCrossRefGoogle Scholar
  169. 169.
    Johansen, J.S., Williamson, M.K., Rice, J.S. and Price, P.A. (1992) Identification of proteins secreted by human osteoblastic cells in culture. Journal of Bone and Mineral Research 7), 501–512.PubMedGoogle Scholar
  170. 170.
    Jukkola, A., Risteli, L., Melkko, J. and Risteli, J. (1993) Procollagen synthesis and extracellular matrix deposition in MG-63 osteosarcoma cells. Journal of Bone and Mineral Research 8), 651–657.PubMedGoogle Scholar
  171. 171.
    Ali, N.N., Rowe, J. and Teich, N.M. (1996) Constitutive expression of non-bone/liver/kidney alkaline phosphatase in human osteosarcoma cell lines. Journal of Bone and Mineral Research 11), 512–520.PubMedGoogle Scholar
  172. 172.
    Hassager, C., Fitzpatrick, L.A., Spencer, E.M. et al. (1992) Basal and regulated secretion of insulin-like growth factor binding proteins in osteoblast-like cells is cell line specific. Journal of Clinical Endocrinology and Metabolism 75), 228–233.PubMedCrossRefGoogle Scholar
  173. 173.
    Okazaki, R., Conover, CA., Harris, S.A. et al. (1995) Normal human osteoblast-like cells consistently express genes for insulin-like growth factors I and II but transformed human osteoblast cell lines do not. Journal of Bone and Mineral Research 10), 788–795.PubMedGoogle Scholar
  174. 174.
    Keeting. P.E., Scott, R.E., Colvard, D.S. et al. (1992) Development and characterization of a rapidly proliferating, well-differentiated cell line derived from normal adult human osteoblast-like cells transfected with SV40 large T antigen. Journal of Bone and Mineral Research 7), 127–136.PubMedCrossRefGoogle Scholar
  175. 175.
    Harris, S.A., Enger, R.J., Riggs, B.L. and Speisberg, T.C. (1995) Development and characterization of a conditionally immortalized human fetal osteoblastic cell line. Journal of Bone and Mineral Research 10), 178–186.PubMedGoogle Scholar
  176. 176.
    Walker, K.E., Houghton, A., Russell, R.G.G. and Stringer, B.M.J. (1995) Identification of conditionally immortalised human osteoprogenitor cell lines responsive to estrogen. Bone 17), 565 (Abstr. 30).CrossRefGoogle Scholar
  177. 177.
    Fisher, L.W., Gehron Robey, P., Tuross, N. et al. (1987) The 24 000 Mr phosphoprotein from developing bone is the N-propeptide of the alphal chain of type I collagen. Journal of Biological Chemistry 262), 13457–13463.PubMedGoogle Scholar
  178. 178.
    Prince, P.A., Ponthermore, J.G. and Deftos, LJ. (1980) A new biochemical marker for bone metabolism. Journal of Clinical Investigation 66), 878–883.CrossRefGoogle Scholar
  179. 179.
    Fisher, L.W., Stubbs, J.T. III and Young, M.F. (1995) Antisera and cDNA probes to human and certain animal model bone matrix noncollagenous proteins. Acta Orthopedica Scandinavica (Suppl. 266)66), 61–65.Google Scholar
  180. 180.
    Bancroft, J.D. (1990) Enzyme histochemistry, in Theory and Practice of Histological Techniques, 3rd edn, (eds J.D. Bancroft and A. Stevens), Churchill Livingstone, London, pp. 379–399.Google Scholar

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© Chapman and Hall Ltd 1998

Authors and Affiliations

  • Francis J. Hughes
  • Jane E. Aubin

There are no affiliations available

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