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Establishment of an osteogenic cell line derived from adult mouse bone marrow stroma by use of a recombinant retrovirus

  • Molecular and Cellular Biology
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Summary

In order to characterize fibroblastic colonyforming units (CFU-F) from murine bone marrow in relation to osteogenesis, adherent cells of 7-day-old BALB/c mouse bone marrow cultures were infected with a recombinant retrovirus (N2/δfosB) containing the bacterial neomycin resistance gene. One of the G418-resistant clones, MN7, was selected for further analysis on the basis of its high expression of the bone-specific alkaline phosphatase. The cells have now been in culture for more than 1 year and maintain a stable phenotype. The osteogenic nature of the immortalized clone MN7 was demonstrated as follows: (1) Mineralization was detected by 85Sr uptake and with the Von Kossa staining method only after in vitro cultivation on a collagen type I matrix. (2) Osteoblastic phenotype markers, including the synthesis of type I collagen, osteonectin, and the bonespecific isoenzyme of alkaline phosphatase were expressed in vitro. (3) MN7 cells responded to bone effectors such as parathyroid hormone and 1,25-dihydroxyvitamin D3. (4) Intraperitoneal injection of MN7 cells into 1-day-old BALB/c mice produced typical osteosarcomas in all animals. We conclude that MN7, derived entirely in vitro from a stromal CFU-F colony, represents a stable murine osteosarcoma cell line expressing the osteoblastic phenotype and provides the first direct evidence needed to establish adult mouse marrow-derived, nonhematopoietic stromal cells as osteoprogenitors.

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Authors and Affiliations

  1. Department of Biochemistry, Lab of Biotechnology, University of Antwerp, UIA, B-2610, Wilrijk, Belgium

    Eric Mathieu & Jozef Merregaert

  2. Department of Radioprotection, SCK, B-2400, Mol, Belgium

    Greet Schoeters & Frank Vander Plaetse

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  1. Eric Mathieu
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  2. Greet Schoeters
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  3. Frank Vander Plaetse
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  4. Jozef Merregaert
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Mathieu, E., Schoeters, G., Vander Plaetse, F. et al. Establishment of an osteogenic cell line derived from adult mouse bone marrow stroma by use of a recombinant retrovirus. Calcif Tissue Int 50, 362–371 (1992). https://doi.org/10.1007/BF00301635

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  • DOI: https://doi.org/10.1007/BF00301635

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