Mineralization in vitro of matrix formed by osteoblasts isolated by collagenase digestion

doi:10.1111/j.1432-0436.1985.tb00310.x

Differentiation

Volume 29, Issue 2, July 1985, Pages 160-168

https://doi.org/10.1111/j.1432-0436.1985.tb00310.x Get rights and content

Abstract

Osteoblasts from calvaria of 18-day-old fetal Sprague-Dawley rats were isolated using a dissecting procedure followed by collagenase digestion. Freshly isolated or previously frozen cells were cultured for up to 4 weeks in a Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum and 50 μg/ml ascorbic acid, with or without 10 mM β-glycerophosphate. Most of the cells were alkaline phosphatase positive throughout the culture period and expressed a type-I collagen as assessed by immunofluorescence. Cells cultured in the presence of β-glycerophosphate formed a matrix with type-I collagen in 7 days. The matrix underwent mineralization in less than 2 weeks. In the absence of β-glycerophosphate, only the formation of a nonmineralized matrix was observed. Electron-microscopic examination revealed osteoblasts embedded in a dense network of collagen fibers, with a well-defined mineralization process in association with matrix vesicles. Scanning electron-microscopy showed that the matrix composed of layers of irregularly shaped spread cells with smooth surfaces trapped in a fiber matrix. No mineralization process was observed when rat skin fibroblasts were cultured under similar conditions. These data demonstrate the ability of enzymatically isolated osteoblasts cultured in the presence of β-glycerophosphate to form bone in vitro, and that this process is similar to bone formation in vivo.

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Mineralization in vitro of matrix formed by osteoblasts isolated by collagenase digestion

Abstract

Dev Biol

Dev Biol

FEBS Lett

Differentiation

Arch Biochem Biophys

Techniques for the preservation of three dimensional structure in preparing specimens for electron microscope

Trans NY Acad Sci

Isolation of bone cell clones with differences in growth, hormone responses and extracellular matrix production

J Cell Biol

Formation of bone tissue in culture from isolated bone cells

J Cell Biol

Current concepts of the physiology and biochemistry of calcification. III. Basic science and pathology

Clin Orthop

Osteoblasts isolated from mouse calvaria initiate matrix mineralization in culture

J Cell Biol

Collagen immunotyping in human liver: light and electron microscope study

J Histochem Cytochcm

Establishment of a clonal osteogenic cell line from newborn mouse calvaria

Jpn J Oral Biol

The migration of osteoblasts

Cell Tissue Res

Biochemical characterization with parathormone and calcitonin of isolated bone cells: provisional identification of osteoclasts and osteoblasts

Endocrinology

Bone formation and calcification by isolated osteoblast-like cells

J Cell Biol