Skip to main content
SpringerLink
Log in

Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

Summary

Cells from fetal or neonatal skeleton can synthesize bone-like tissue in vitro. In contrast, formation of bone-like tissue in vitro by cells derived from adult animals has rarely been reported and has not been achieved using cells from bone marrow. We have explored development of bone-like tissue in vitro by bone marrow stromal cells. Marrow stromal cells obtained from 40–43-day-old Wistar rats were grown in primary culture for 7 days and then subcultured for 20–30 days. Cells were cultured in either α-minimal essential medium containing 15% fetal bovine serum, antibiotics, and 50 μg/ml ascorbic acid, or the above medium supplemented with either 10 mM Na-β-glycerophosphate, 10-8 M dexamethasone, or a combination of both. Cultures were examined using phase-contrast microscopy, undemineralized and demineralized tissue histology, histochemistry (for alkaline phosphatase activity), immunohistochemistry (for collagen type, osteonectin, and bone Glaprotein), scanning and transmission electron microscopy, energy dispersive X-ray microanalysis, and X-ray diffraction. Collagenous, mineralized nodules exhibiting morphological and ultrastructural characteristics similar to bone were formed in the cultures, but only in the presence of both β-glycerophosphate and dexamethasone. Cells associated with the nodules exhibited alkaline phosphatase activity. The matrix of the nodules was composed predominantly of type-I collagen and both osteonectin and Glaprotein were present. X-ray microanalysis showed the presence of Ca and P, and X-ray diffraction indicated the mineral to be hydroxyapatite. The nodules were also examined for bone morphogenetic protein-like activity. Paired diffusion chambers containing partly demineralized nodules and fetal muscle were implanted intraperitonealy in rats. Induction of cartilage in relation to muscle was observed histologically after 40 days in the chambers. This finding provided further support for the bone-like nature of the nodules. The observations show that bone-like tissue can be synthesized in vitro by cells cultured from young-adult bone marrow, provided that the medium contains both β-glycerophosphate and, particularly, dexamethasone.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Anderson RE, Kemp JW, Jee WSS, Woodbury DM (1984a) Ion-transporting ATP-ases and matrix mineralization in cultured osteoblast-like cells. In Vitro 20:837–846

    Google Scholar 

  • Anderson RE, Kemp JW, Jee WSS, Woodbury DM (1984b) Effects of cortisol and fluoride on ion-transporting ATPase activities in cultured osteoblast-like cells. In Vitro 20:847–855

    Google Scholar 

  • Ashton BA, Allen TD, Howlett CR, Eaglesom CC, Hattori A, Owen M (1980) Formation of bone and cartilage by marrow stromal cells in diffusion chambers in vivo. Clin Orthop 151:294–307

    Google Scholar 

  • Aubin JE, Heersche JNM, Merrilees MJ, Sodek J (1982) Isolation of bone cell clones with differences in growth, hormone responses and extracellular matrix production. J Cell Biol 92:452–461

    Google Scholar 

  • Auf'mkolk B, Hauschka PV, Schwartz ER (1985) Characterization of human bone cells in culture. Calcif Tissue Int 37:228–235

    Google Scholar 

  • Barnes MJ (1975) Function of ascorbic acid in collagen metabolism. Ann NY Acad Sci 258:264–277

    Google Scholar 

  • Baron R, Vignery A, Neff L, Silverglate A, Santa Maria A (1983) Processing of undecalcified bone specimens for bone histomorphometry. In: Recker RR (ed) Bone Histomorphometry, Techniques and Interpretation. CRC Press, Boca Raton, pp 3–13

    Google Scholar 

  • Bellows CG, Aubin JE, Heersche JNM, Antosz NM (1986) Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations. Calcif Tissue Int 38:143–154

    Google Scholar 

  • Bellows CG, Aubin JE, Heersche JNM (1987) Physiological concentrations of glucocorticoids stimulate formation of bone nodules from isolated rat calvaria cells in vitro. Endocrinology 121:1985–1992

    Google Scholar 

  • Binderman I, Duksin D, Harell A, Katzir (Katchalski) E, Sachs L (1974) Formation of bone tissue in culture from isolated mesenchymal cells. J Cell Biol 61: 427–439

    Google Scholar 

  • Bornstein P, Sage H (1980) Structurally distinct collagen types. Ann Rev Biochem 49:957–1003

    Google Scholar 

  • Boskey AL (1981) Current contents of the physiology and biochemistry of calcification. Clin Orthop 157:225–257

    Google Scholar 

  • Boyde A (1972) Scanning electron microscope studies of bone. In: Bourne GH (ed) Biochemistry and Physiology of Bone. Vol. 1, 2nd ed. Academic Press, New York, pp 259–310

    Google Scholar 

  • Bronckers ALJJ, Gay S, DiMuzio MT, Butler WT (1985) Immuno-localization of γ-carboxyglutamic acid containing proteins in developing rat bones. Coll Rell Res 5:273–281

    Google Scholar 

  • Burger EH, Boonekamp PM, Nijweide PJ (1986) Osteoblast and osteoclast precursors in primary cultures of calvaria bone cells. Anat Rec 214:32–40

    Google Scholar 

  • Burgeson RE (1982) Genetic heterogeneity of collagens. J Invest Dermatol 79:255–305

    Google Scholar 

  • Cameron DA (1972) The ultrastructure of bone. In: Bourne GH (ed) Biochemistry and Physiology of Bone. Vol. 1, 2nd ed. Academic Press, New York, pp 191–236

    Google Scholar 

  • Canalis E (1983) Effect of glucocorticoids on type I collagen synthesis, alkaline phosphatase activity, and deoxyribonucleic acid content in cultured rat calvaria. Endocrinology 112:931–939

    Google Scholar 

  • Caplan AI (1977) Muscle, cartilage and bone development and differentiation from chick limb mesenchymal cells. In: Ede DA, Hinchliffe JR, Balls M (eds) Vertebrate Limb and Somite Morphogenesis. Cambridge University Press, Cambridge, pp 199–213

    Google Scholar 

  • Catherwood BD (1985) 1,25-dihydroxycholecalciferol and glucocorticosteroid regulation of adenylate cyclase in an osteoblastlike cell line. J Biol Chem 260:736–743

    Google Scholar 

  • Cerro M del, Cogen J, Cerro C del (1980) Stevenel's blue, an excellent stain for optical microscopical study of plastic embedded tissues. Microsc Acta 83:117–121

    Google Scholar 

  • Chen TL, Feldman D (1978) Glucocorticoid potentiation of the adenosine 3′,5′-monophosphate response to parathyroid hormone in cultured rat bone cells. Endocrinology 102:589–596

    Google Scholar 

  • Chen TL, Feldman D (1979) Glucocorticoid receptors and action in subpopulations of cultured rat bone cells. J Clin Invest 63:750–758

    Google Scholar 

  • Chen TL, Aronow L, Feldman D (1977) Glucocorticoid receptors and inhibition of bone cell growth in primary culture. Endocrinology 100:619–628

    Google Scholar 

  • Chen TL, Cone CM, Feldman D (1983) Glucocorticoid modulation of cell proliferation in cultured osteoblast-like bone cells: Differences between rat and mouse. Endocrinology 112:1739–1745

    Google Scholar 

  • Choe J, Stern P, Feldman D (1978) Receptor mediated glucocorticoid inhibition of protein synthesis in isolated bone cells. J Steroid Biochem 9:265–271

    Google Scholar 

  • Chyun YS, Kream BE, Raisz LG (1983) Cortisol decreases bone formation by inhibiting periosteal cell proliferation. Endocrinology 114:477–480

    Google Scholar 

  • Delmas PD, Tracy RP, Riggs BL, Mann KG (1984) Identification of the non-collagenous proteins of bovine bone by two-dimensional gel electrophoresis. Calcif Tissue Int 36:308–316

    Google Scholar 

  • Dietrich JW, Canalis EM, Maina DM, Raisz LG (1979) Effects of glucocorticoids on fetal rat bone collagen synthesis in vitro. Endocrinology 104:715–721

    Google Scholar 

  • Drury RAB, Wallington EA (1967) E. A. Carleton's Histological technique. 4th ed. Oxford University Press, New York, pp 151, and pp 242–245

    Google Scholar 

  • Ecarot-Charrier B, Glorieux FH, van der Rest M, Pereira G (1983) Osteoblasts isolated from mouse calvaria initiate matrix mineralization in culture. J Cell Biol 96:639–643

    Google Scholar 

  • Epstein EH Jr, Munderloh NH (1975) Isolation and characterization of CNBr peptides of human [α1 (III)]3 collagen and tissue distribution of [α1(I)]2 and [α1(III)]3 collagens. J Biol Chem 250:9304–9312

    Google Scholar 

  • Feldman D, Dziak R, Koehler R, Stern P (1975) Cytoplasmic glucocorticoid binding proteins in bone cells. Endocrinology 96:29–36

    Google Scholar 

  • Friedenstein AJ (1973) Determined and inducible osteogenic precursor cells. In: Hard tissue growth, repair and remineralization. Ciba Foundation Symposium II (new series). Elsevier/ Excerpta Medica/North-Holland Assoc. Scientific Publ., Amsterdam, pp 169–185

    Google Scholar 

  • Friedenstein AJ (1976) Precursor cells of mechanocytes. Int RevCytol 43:327–355

    Google Scholar 

  • Friedenstein AJ, Chaylakhjan RK, Lalykina KS (1970) The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells. Cell Tissue Kinet 3:393–403

    Google Scholar 

  • Friedenstein AJ, Chailaklyan RK, Latsinik NV, Panasyuk AF, Keiliss-Borok IV (1974) Stromal cells responsible for transferring the microenvironment of hemopoetic tissues. Transplantation 17:331–340

    Google Scholar 

  • Gehron Robey P, Termine JD (1985) Human bone cells in vitro. Calcif Tissue Int 37:453–460

    Google Scholar 

  • Gehron Robey P, Beresford JN, Naglak DM, Heywood BR (1986) Demonstration of osteogenesis in vitro. J Cell Biol 103: No. 384 [Abstr]

  • Gennari C (1985) Glucocorticoids and bone. In: Peck WA (ed) Bone and mineral research/3. Elsevier, Amsterdam, pp 213–231

    Google Scholar 

  • Green S, Chambon P (1986) A superfamily of potentially oncogenic hormone receptors. Nature 324:615–617

    Google Scholar 

  • Hahn TJ, Westbrook SL, Halstead LR (1984) Cortisol modulation of osteoblast metabolic activity in cultured neonatal rat bone. Endocrinology 114:1864–1870

    Google Scholar 

  • Ham AW (1974) Bone. In: Ham AW (ed) Histology. 7th edition. J.B. Lippincott Co., Philadelphia, pp 378–447

    Google Scholar 

  • Harell A, Binderman I, Guez M (1976) Tissue culture of bone cells: Mineral transport, calcification and hormonal effects. Israel J Med Sci 12:115–123

    Google Scholar 

  • Hauschka PV (1985) Osteocalcin and its functional domains. In: Butler WT (ed) The chemistry and biology of mineralized tissues. EBSCO Media, Birmingham, Alabama, pp 149–158

    Google Scholar 

  • Haynes RC Jr, Murad F (1985) Adrenocorticotropic hormone; adrenocortical steroids and their synthetic analogs; inhibitors of adrenocortical steroid biosynthesis. In: Goodman Gilman A, Goodman LS, Rall TW, Murad F (eds) Goodman and Gilman's. The pharmacological basis of therapeutics. 7 ed. MacMillan Publishing Co., New York, pp 1459–1489

    Google Scholar 

  • Hirano H, Urist MR (1981) Bone-forming and bone-resorbing cell lines derived from bone marrow in tissue culture. Clin Orthop 154:234–248

    Google Scholar 

  • Howlett CR, Owen M, Cave J, Williamson M, Bab I, Maybee S, Trifitt JT (1984) In vitro mineralization and alkaline phosphatase activity in cultures of rabbit bone marrow stromal cells. Calcif Tissue Int 36 [Suppl 2]: S 67

    Google Scholar 

  • Howlett CR, Cave J, Williamson M, Farmer J, Ali SY, Bab I, Owen ME (1986) Mineralization in in vitro cultures of rabbit marrow stromal cells. Clin Orthop 213:251–263

    Google Scholar 

  • Jande SS (1971) Fine structural study of osteocytes and their surrounding bone matrix with respect to their age in young chicks. J Ultrastruc Res 37:279–300

    Google Scholar 

  • Jee WSS, Blackwood EL, Dockum NL, Haslam RK, Kircl FA (1966) Bio-assay of responses of growing bones to cortisol. Clin Orthop 49:39–63

    Google Scholar 

  • Jett S, Wu K, Duncan H, Frost HM (1970) Adrenalcorticosteroid and salicylate actions on human and canine haversian bone formation and resorption. Clin Orthop 68:301–315

    Google Scholar 

  • Johnson GD, Davidson RS, McNamee RC, Russel G, Goodwin D, Holborrow EJ (1982) Fading of immunofluorescence during microscopy: A study of the problem and its remedy. J Immunol Methods 55:231–242

    Google Scholar 

  • Kodicek E (1965) The effect of ascorbic acid on biosynthesis of components of connective tissue. In: Fitton Jackson S, Harkness RD, Patridge SM, Tristram GR (eds) Structure and Function of Connective and Skeletal Tissue. Butterworths, London, pp 307–319

    Google Scholar 

  • Koshihara Y, Kawamura M, Oda H, Higaki S (1987) In vitro calcification in human osteoblastic cell line. Biochem Biophys Res Commun 145:651–657

    Google Scholar 

  • Lian JB, Friedman PA (1978) The vitamin K dependent synthesis of γ-carboxyglutamic acid by bone microsomes. J Biol Chem 253:6623–6626

    Google Scholar 

  • Luk SC, Nopajaroonski C, Simon GT (1974) The ultrastructure of cortical bone in young adult rabbits. J Ultrastruc Res 46:184–205

    Google Scholar 

  • Luria EA, Owen ME, Friedenstein AJ, Morris JF, Kuznetsow SA (1987) Bone formation in organ cultures of bone marrow. Cell Tissue Res 248:449–454

    Google Scholar 

  • Maniatopoulos C, Melcher AH (1988) Parameters affecting bone-like tissue formation in vitro by bone marrow stromal cells. J Dent Res 67:290 [IADR Abstr. No 1418]

    Google Scholar 

  • Manolagas SC, Anderson DC (1979) Glucocorticoids regulate the concentration of 1,25-dihydroxycholecalciferol receptors in bone. Nature 277:314–315

    Google Scholar 

  • Marshall S, Huang HH, Kledzik GS, Campbell GA, Meites J (1978) Glucocorticoid regulation of prolactin receptors in kidneys and adrenals of male rats. Endocrinology 102:869–875

    Google Scholar 

  • McCulloch CAG, Tenenbaum HC (1986) Dexamethasone induces proliferation and terminal differentiation of osteogenic cells in tissue culture. Anat Rec 215:397–402

    Google Scholar 

  • McLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative: A new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083

    CAS  PubMed  Google Scholar 

  • Miller EJ (1973) A Review of biochemical studies on the genetically distinct collagens of the skeletal system. Clin Orthop 92:260–280

    Google Scholar 

  • Nefussi J-R, Boy-Lefevre ML, Boulekbache H, Forest N (1985) Mineralization in vitro of matrix formed by osteoblasts isolated by collagenase digestion. Differentiation 29:160–168

    Google Scholar 

  • Nishimoto SK, Price PA (1979) Proof that the γ-carboxyglutamic acid-containing protein is synthesized in calf bone: comparative synthesis rate and effect of coumadin on synthesis. J Biol Chem 254:437–441

    Google Scholar 

  • Nishimoto SK, Price PA (1980) Secretion of the vitamin K-dependent protein of bone by rat osteosarcoma cells. J Biol Chem 25:6579–6583

    Google Scholar 

  • Osdoby P, Caplan AI (1976) The possible differentiation of osteogenic elements in vitro, from chick limb mesodermal cells. Dev Biol 52:283–299

    Google Scholar 

  • Osdoby P, Caplan AI (1979) Osteogenesis in cultures of limb mesenchymal cells. Dev Biol 73:84–102

    Google Scholar 

  • Osdoby P, Caplan AI (1980) A scanning electron microscopic investigation of in vitro osteogenesis. Calcif Tissue Int 30:43–50

    Google Scholar 

  • Otsuka K, Yao K-L, Wasi S, Tung PS, Aubin JE, Sodek J (1984) Biosynthesis of Osteonectin by fetal porcine calvaria cells in vitro. J Biol Chem 259:9805–9812

    Google Scholar 

  • Owen M (1985) Lineage of osteogenic cells and their relationship to the stromal system. In: Peck WA (ed) Bone and Mineral Research/3. Elsevier, Amsterdam, pp 1–25

    Google Scholar 

  • Owen M, Howlett R, Maybee S, Cave J, Triffitt JT (1984) Expression of alkaline phosphatase activity in in vitro cultures of marrow stromal cells. Calcif Tissue Int 36:467 (Abstr)

    Google Scholar 

  • Pearse AGE (1972) Histochemistry, Theoretical and Applied. Vol. 2, 3rd ed. Churchill, Livingston, pp 1138, 1405

    Google Scholar 

  • Peck WA, Brandt W, Miller I (1967) Hydrocortisone-induced inhibition of protein synthesis and uridine incorporation in isolated bone cells in vitro. Proc Natl Acad Sci USA 57:1599–1606

    Google Scholar 

  • Posner AS (1985) The mineral of bone. Clin Orthop 200:87–99

    Google Scholar 

  • Price PA, Baukol SA (1980) 1,25-dihydroxyvitamin D3 increases synthesis of the vitamin K-dependent bone protein by osteosarcoma cells. J Biol Chem 255:11660–11663

    Google Scholar 

  • Price PA, Williamson MK, Lothringer JW (1981) Origin of the vitamin K-dependent bone protein found in plasma and its clearance by kidney and bone. J Biol Chem 256:12760–12766

    Google Scholar 

  • Price PJ, Gregory EA (1982) Relationship between in vitro growth promotion and biophysical and biochemical properties of the serum supplement. In Vitro 18:576–584

    Google Scholar 

  • Rao LV, Kalliecharan R, Wang H-M, Heersche JNM, Sodek J (1979) Specific immunohistochemical localization of type I collagen in porcine periodontal tissue using the peroxidase-labelled antibody technique. Histochem J 11:73–82

    Google Scholar 

  • Rath NC, Reddi AH (1979) Influence of adrenalectomy and dexa-methasone on matrix-induced endochondral bone differentiation. Endocrinology 104:1698–1703

    Google Scholar 

  • Ries WL, Gong JK, Sholley MM (1985) A comparative study of osteoblasts in situ versus isolated specimend. Am J Anat 172:57–73

    Google Scholar 

  • Rodan GA, Rodan SB (1983) Expression of the osteoblastic phenotype. In: Peck WA (ed) Bone and Mineral Research. Annual 2. Elsevier, Amsterdam, pp 244–285

    Google Scholar 

  • Rosenberg L (1971) Chemical basis for the histological use of safranin O in the study of articular cartilage. J Bone Joint Surg 53A: 69–82

    Google Scholar 

  • Sampath TK, Reddi AH (1981) Dissociative extraction and reconstitution of extracellular matrix components involved in local bone differentiation. Proc Natl Acad Sci USA 78:7599–7603

    Google Scholar 

  • Sampath TK, Muthukumaran N, Reddi AH (1987) Isolation of osteogenin, an extracellular matrix-associated, bone-inductive protein, by heparin affinity chromatography. Proc Natl Acad Sci USA 84:7109–7113

    Google Scholar 

  • Scott DM, Kent GN, Cohn DV (1980) Collagen synthesis in cultured osteoblast-like cells. Arch Biochem Biophys 201:384–391

    Google Scholar 

  • Seyedin SM, Thomas TC, Thompson AY, Rose DM, Piez KA (1985) Purification and characterization of two cartilage-inducing factors from bovine demineralized bone. Proc Natl Acad Sci USA 82:2267–2271

    Google Scholar 

  • Simmons DJ, Kunin AS (1967) Autoradiographic and biochemical investigation of the effect of cortisone on the bones of the rat. Clin Orthop 55:201–215

    Google Scholar 

  • Stenner DD, Tracy RP, Riggs BL, Mann KG (1986) Human platelets contain osteonectin, a major protein of mineralized bone. Proc Natl Acad Sci 83:6892–6896

    Google Scholar 

  • Sudo H, Kodama HA, Amagai Y, Yamamoto S, Kasai SC (1983) In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria. J Cell Biol 96:191–198

    Google Scholar 

  • Syftestad GT, Lucas PA, Caplan AI (1985) The in vitro chondrogenic response of limb-bud mesenchyme to a water-soluble fraction prepared from demineralized bone matrix. Differentiation 29:230–237

    Google Scholar 

  • Takaoka K, Yoshikawa H, Shimizu N, Ono K, Amitani K, Nakata Y, Sakamoto Y (1981) Purification of a bone-inducing substance (osteogenic factor) from a murine osteosarcoma. Biomed Res 2:466–471

    Google Scholar 

  • Tashjian AH Jr, Osborne R, Maina D, Knaian A (1977) Hydrocortisone increases the number of receptors for thyrotropin releasing hormone on pituitary cells in culture. Biochem Biophys Res Commun 79:333–340

    Google Scholar 

  • Tenenbaum HC (1981) Role of organic phosphate in mineralization of bone in vitro. J Dent Res 60 [Sp. Iss. C]: 1586–1589

    Google Scholar 

  • Tenenbaum HC, Heersche JNM (1982) Differentiation of osteoblasts and formation of mineralized bone in vitro. Calcif Tissue Int 34:76–79

    Google Scholar 

  • Tenenbaum HC, Heersche JNM (1985) Dexamethasone stimulates osteogenesis in chick periosteum in vitro. Endocrinology 117:2211–2217

    Google Scholar 

  • Termine JD (1983) Osteonectin and other newly described proteins of developing bone. In: Peck WA (ed) Bone and Mineral Research Annual 1: A Yearly Survey of Developments in the Field of Bone and Mineral Metabolism. Expecta Medica, Amsterdam, pp 144–156

    Google Scholar 

  • Termine JD, Belcourt AB, Conn KM, Kleinman HK (1981a) Mineral and collagen-binding proteins of fetal calf bone. J Biol Chem 20:10403–10408

    Google Scholar 

  • Termine JD, Kleinman HK, Whitson SW, Conn KM, McGarvey ML, Martin GR (1981b) Osteonectin, a bone-specific protein linking mineral to collagen. Cell 26:99–105

    Article  CAS  PubMed  Google Scholar 

  • Tibone KW, Bernard GW (1982) A new in vitro model of intra-membranous osteogenesis from adult bone marrow stem cells. In: Dixon AD, Sarnat BG (eds) Factors and Mechanisms Influencing Bone Growth. Alan R. Liss Inc., New York, pp 107–123

    Google Scholar 

  • Tsuji T, McCulloch CAG, Melcher AH (1988) Effect of age on osteogenesis in vitro. J Dent Res 67:234 (IADR abstr. 970)

    Google Scholar 

  • Tsuru S, Kitani H, Oguchi M, Mashiko M, Zinnaka Y, Shimomura Y (1984) Separation of osteoblast-like cells from bone marrow by fluorescence-activated cell sorting. J Histochem Cytochem 32:43–48

    Google Scholar 

  • Tung PS, Domenicucci C, Wasi S, Sodek J (1985) Specific immuno-histochemical localization of osteonectin and collagen types I and III in fetal and adult porcine dental tissues. J Histochem Cytochem 33:531–540

    Google Scholar 

  • Urist MR (1980) Heterotopic bone formation. In: Urist MR (ed) Fundamental and Clinical Bone Physiology. J.B. Lippincott Co., Philadelphia, pp 369–392

    Google Scholar 

  • Urist MR, Mizutani H, Conover MA, Lietze A, Finerman GAM (1982) Dentin, bone, and osteosarcoma tissue bone morphogenetic proteins. In: Dixon AD, Sarnat BG (eds) Factors and Mechanisms Influencing Bone Growth. Alan R. Liss Inc., New York, pp 61–81

    Google Scholar 

  • Urist MR, Hou YK, Brownell AG, Holh WM, Buyske J, Lietze A, Tempst P, Hunkapiller M, Delange RJ (1984) Purification of bovine bone morphogenetic protein by hydroxyapatite cromatography. Proc Natl Acad Sci USA 81:371–375

    Google Scholar 

  • Wang H-M, Nanda V, Rao LV, Melcher AH, Heersche JNM, Sodek J (1980) Specific immunohistochemical localization of type III collagen in porcine periodontal tissues using the peroxidase-antiperoxidase method. J Histochem Cytochem 28:1215–1223

    Google Scholar 

  • Wasi S, Otsuka K, Yao K-L, Tung PS, Aubin JE, Sodek J, Termine JD (1984) An osteonectin protein in porcine periodontal ligament and its synthesis by periodontal ligament fibroblasts. Can J Biochem Cell Biol 62:470–478

    Google Scholar 

  • Whitson SW, Harrison W, Dunlap MK, Bowers DE Jr, Fisher LW, Gehron Robey P, Termine JD (1984) Fetal bovine bone cells synthesize bone-specific matrix proteins. J Cell Biol 99:607–614

    Google Scholar 

  • Wiestner M, Fischer S, Dessau W, Muller PK (1981) Collagen types synthesized by isolated calvarium cells. Exp Cell Res 133:115–125

    Google Scholar 

  • Williams DC, Boder GB, Toomey RE, Paul DC, Hillman CC Jr, King KL, van Frank RM, Johnson C Jr (1980) Mineralization and metabolic response in serially passaged adult rat bone cells. Calcif Tissue Int 30:233–246

    Google Scholar 

  • Wong GL (1979) Basal activities and hormone responsiveness of osteoclast-like and osteoblast-like bone cells are regulated by glucocorticoids. J Biol Chem 254:6337–6340

    Google Scholar 

  • Zung P, Domenicucci C, Wasi S, Kuwata F, Sodek J (1986) Osteonectin is a minor component of mineralized connective tissues in rat. Biochem Cell Biol 64:356–362

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. M.R.C. Programme in Dental Implantology, Faculty of Dentistry, University of Toronto, Toronto, Ontario, Canada

    C. Maniatopoulos &  A. H. Melcher

  2. M.R.C Group in Periodontal Physiology, Faculty of Dentistry, University of Toronto, Toronto, Ontario, Canada

    J. Sodek

Authors
  1. C. Maniatopoulos
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. J. Sodek
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. H. Melcher
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Maniatopoulos, C., Sodek, J. & Melcher, A.H. Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats. Cell Tissue Res. 254, 317–330 (1988). https://doi.org/10.1007/BF00225804

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00225804

Key words

Search

Navigation