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    Factors that promote progressive development of the osteoblast phenotype in cultured fetal rat calvaria cells

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    Authors
    Aronow, Michael A.
    Gerstenfeld, Louis C.
    Owen, Thomas A.
    Tassinari, Melissa S.
    Stein, Gary S.
    Lian, Jane B.
    UMass Chan Affiliations
    Department of Cell Biology
    Department of Orthopedic Surgery
    Document Type
    Journal Article
    Publication Date
    1990-05-01
    Keywords
    Alkaline Phosphatase
    Animals
    *Bone Development
    Calcium
    Cell Differentiation
    Cells, Cultured
    Chickens
    Collagen
    DNA
    Osteoblasts
    Osteocalcin
    Proteins
    Rats
    Cell Biology
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    Link to Full Text
    http://dx.doi.org/10.1002/jcp.1041430203
    Abstract
    Rat calvaria osteoblasts derived from 21-day-old fetal rat pups undergo a temporal expression of markers of the osteoblast phenotype during a 5 week culture period. Alkaline phosphatase and osteocalcin are sequentially expressed in relation to collagen accumulation and mineralization. This pattern of expression of these osteoblast parameters in cultured rat osteoblasts (ROB) is analogous to that seen in vivo in developing fetal rat calvaria tissue (Yoon et. al: Biochem. Biophis. Res. Commun. 148:1129, 1987) and is similar to that observed in cultures of subcultivated 16-day-old embryonic chick calvaria-derived osteoblasts (COB) (Gerstenfeld, et.al: Dev. Biol. 122:46, 1987). While the cellular organization of subcultivated COB and primary ROB cultures are somewhat different, the temporal expression of the parameters remains. Both the rat and chick culture systems support formation of matrix mineralization even in the absence of beta-glycerol-phosphate. A systematic examination of factors which constitute conditions supporting complete expression of the osteoblast phenotype in ROB cultures indicate requirements for specific serum lots, ascorbic acid and the ordered deposition of mineral in the extracellular matrix. The present studies suggest that formation of a collagenous matrix, dependent on ascorbic acid, is requisite for expression of the osteoblast phenotype. In ROB cultures, expression of osteocalcin synthesis occurs subsequent to initiation of alkaline phosphatase activity and accompanies the formation of mineralized nodules. Thus, extracellular matrix mineralization (deposition of hydroxyapatite) is required for complete development of the osteoblast phenotype, as reflected by a 200-fold increase in osteocalcin synthesis. These data show the temporal expression of the various osteoblast parameters during the formation and mineralization of an extracellular matrix can provide markers reflective of various stages of osteoblast differentiation/maturation in vitro.
    Source
    J Cell Physiol. 1990 May;143(2):213-21. Link to article on publisher's site
    DOI
    10.1002/jcp.1041430203
    Permanent Link to this Item
    http://hdl.handle.net/20.500.14038/49472
    PubMed ID
    2332447
    Related Resources
    Link to Article in PubMed
    ae974a485f413a2113503eed53cd6c53
    10.1002/jcp.1041430203
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