Summary
The process of mineralization in cells prepared either by neutral protease digestion (Pro I) or by collagenase digestion (fifth cycle, Col V) from fetal rat mandible was studied in vitro. Alkaline phosphatase (ALPase) activity of cells in Pro I was low on day 3, increased rapidly from day 8, and reached a maximum on day 16, whereas that in Col V was high on day 2, then declined and thereafter elevated to reach a maximum on day 13. Both cell populations synthesized type I collagen in cell matrix and medium. Type III collagen was observed in cell matrix of Pro I on day 14 and 21. There was α2 band of type V collagen in cell matrix of Pro I on day 21. Calcium deposition could be detected from day 14 in Pro I and from day 19 in Col V. The von Kossapositive nodules were found on day 17 in Pro I and day 21 in Col V, respectively. The extracellular matrix in Pro I electron-microscopically consisted of well-banded collagen fibrils with a large number of calcified spherules. An elevation of ALPase activity, collagen synthesis, and mineral deposition occurred sequentially with a time lapse in Col V, and almost simultaneously in Pro I. The number of mineralized nodules was correlated with the density of plated cells in Pro I, but not in Col V. Dexamethasone caused an increase in the number of mineralized nodules in Pro I, but not in Col V, suggesting that Pro I contained osteoprogenitor cells. Thus, the mode of mineralization in cells derived from the mandible may differ depending on the presence of osteoprogenitor cells.
Similar content being viewed by others
References
Nijwide PJ, van Iperen-van Gent AS, Kawilarang-de Haas EWM, van der Plas A, Wassenaar AM (1982) Bone formation and calcification by isolated osteoblast-like cells. J Cell Biol 93:318–323
Sudo H, Kodama H, Amagai Y, Yamamoto S, Kasai S (1983) In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria. J Cell Biol 96: 191–198
Bellows CG, Aubin JE, Heersche JNM, Antosz ME (1986) Mineralized bone nodules formed in vitro from enzymatically released rat calvaria cell populations. Calcif Tissue Int 38:143–154
Gerstenfeld LC, Chipman SD, Glowacki J, Lian JB (1987) Expression of differentiated function by mineralizing cultures of chicken osteoblasts. Dev Biol 122:49–60
Masquelier D, Herbert B, Hauser N, Mermillod P, Schonne E, Remacle C (1990) Morphologic characterization of osteoblastlike cell cultures isolated from newborn rat calvaria. Calcif Tissue Int 47:92–104
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
Wiestner M, Fischer S, Dessau W, Muller PK (1981) Collagen types synthesized by calvarium cells. Exp Cell Res 133:115–125
Aronow MA, Gerstenfeld LC, Owen TA, Tassinari MS, Stein GS, and Lian JB (1990) Factors that prompt progressive development of the osteoblast phenotype in cultured fetal rat calvaria cells. J Cell Physiol 143:213–221
Tenenbaum HC, Limeback H, McCulloch CAG, Hamujee H, Sukhu B, Torontali M (1992) Osteogenic phase-specific coregulation of collagen synthesis and mineralization by β-glycerophosphate in chick periosteal cultures. Bone 13:129–138
Stein GS, Lian JB, Owen TA (1990) Relationship of cell growth to the regulation of tissue-specific gene expression during osteoblast differentiation. FASEB J 4:3111–3123
Collin P, Nefussi JR, Wetterwald A, Nicolas V, Boy-Lefevre M, Fleisch H, Forest N (1992) Expression of collagen, osteocalcin, bone alkaline phosphatase in a mineralizing rat osteoblastic cell culture. Calcif Tissue Int 50:175–183
Owen M, Ashton B (1986) Osteogenic differentiation of skeletal cell populations. In: Ali Y (ed) Cell-mediated calcification and matrix vesicles. Elsevier, Amsterdam, pp 279–284
Owen ME, Cave J, Joyner CJ (1987) Clonal analysis in vitro of osteogenic differentiation of marrow CFU-F. J Cell Sci 87:731–738
McCulloch CAG, Fair CA, Tenebaum HC, Limeback H, Homareau R (1990) Clonal distribution of osteoprogenitor cells in cultured chick periostea: functional relationship to bone formation. Dev Biol 140:352–361
Bellows CG, Aubin JE (1989) Determination of numbers of osteoprogenitor cells present in isolated fetal rat calvaria cells in vitro. Dev Biol 133:8–13
Maniatopoulos C, Sodek J, Melcher AH (1988) Bone formation in vitro by stromal cells obtained from bone marrow of young adult rats. Cell Tissue Res 254:317–330
Labarca C, Paigen K (1980) A simple, rapid and sensitive DNA assay procedure. Anal Biochem 102:344–352
Lowry OH (1955) Micromethods for the assay of enzyme. II. Specific procedures. Alkaline phosphatase. Methods Enzymol 4:371–372
Lowry OH, Rosenbrough NJ, Faar AL (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
Connerty HV, Briggs AR (1966) Determination of serum calcium by means of orthocresolphtalein complexone. Am J Pathol 45:290–296
Brumenkrantz N, Asboe-hansen G (1973) A quick and specific assay of hydroxyproline. Anal Biochem 55:288–291
Koshihara Y, Kawamura M, Endo S, Tsutsumi C, Kodama H, Higaki S (1989) Establishment of human osteoblastic cells derived from periosteum in culture. In Vitro 25:37–43
Sykes B, Piddle B, Francis M, Smith R (1976) The estimation of two collagens from human dermis by interrupted gel electrophoresis. Biochem Biophys Res Commun 72:1472–1480
Zimmermann B, Wachtel HC, Noppe C (1991) Patterns of mineralization in vitro. Cell Tissue Res 263:483–493
Owen TA, Aronow M, Shalhoub V, Barone LM, Wilming L, Tassinari MS, Kennedy MB, Pockwinse S, Lian JB, Stein GS (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. J Cell Physiol 143:420–430
Lomri A, Tran MPV, Hott M (1988) Characterization of endosteal osteoblastic cells isolated from mouse caudal vertebrae. Bone 9:165–175
Daniel JC, Pauli BU, Kuettner KE (1984) Synthesis of cartilage matrix by mammalian chondrocytes in vitro. III. Effects of ascorbate. J Cell Biol 99:1960–1969
Grigoriadis AE, Heersche JNM, Aubin JE (1988) 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
Renny TF, Bhanumathi SI (1992) Relationship between collagen synthesis and expression of the osteoblast phenotype in MC3T3-E1 cells. J Bone Miner Res 7:235–246
Turksen K, Aubin JE (1991) Positive and negative immunoselection of two classes of osteoprogenitor cells. J Cell Biol 114: 373–384
Bellows CG, Heersche JNM, Aubin JE (1989) Effects of dexamethasone on expression and maintenance of cartilage in serum-containing cultures of calvarial cells. Cell Tissue Res 256: 145–151
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Abe, Y., Akamine, A., Aida, Y. et al. Differentiation and mineralization in osteogenic precursor cells derived from fetal rat mandibular bone. Calcif Tissue Int 52, 365–371 (1993). https://doi.org/10.1007/BF00310201
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00310201