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In vitro analysis of bone phenotypes in Col1a1 and Jagged1 mutant mice using a standardized osteoblast cell culture system

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Abstract

The mouse is a valuable model organism for studying bone biology and for unravelling pathological processes in skeletal disorders. In vivo methods like X-ray analysis, DXA measurements, pQCT and μCT are available to investigate the bone phenotype of mutant mice. However, the descriptive nature of such methods does not provide insights into the cellular and molecular bases of the observed bone alterations. Thus, first-line investigations might be complemented by cell culture-based methods to characterize the pathological processes at the cellular level independent from systemic influences. By combining well-established assays, we designed a comprehensive test system to investigate the cellular and molecular phenotype of primary calvarial osteoblasts in mutant mice compared to wild-type controls as a first-line phenotyping method. The compilation of 9 different quantifiable assays allows assessment of general properties of cell growth and investigation of bone-specific parameters at the functional, protein and RNA level in a kinetic fashion throughout a 3-week culture period, thus maximizing the chance to discover and explain new phenotypes in mutant mice. By analyzing mutant mouse lines for Col1a1 and Jag1 (Delta-Notch pathway) that both showed clear alterations in several bone-related parameters we could demonstrate the usefulness of our cell culture system to discriminate between primary (Col1a1) and secondary effects (Jag1) in osteoblasts.

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Acknowledgments

The authors would like to thank Reinhard Seeliger, Susanne Wittich and Michael Schulz for their excellent technical assistance and Dr. Uwe Kornak for critical reading of the manuscript and helpful discussions. This work was supported by NGFN + grant 01GS0850 from the German Federal Ministry of Education and Research.

Conflict of interest

All authors have no conflicts of interest.

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Author notes

  1. Frank Thiele

    Present address: Technische Universitaet Muenchen, Institute of Virology, Klinikum Rechts der Isar, Schneckenburgerstr. 8, 81675, Muenchen, Germany

Authors and Affiliations

  1. Institute of Experimental Genetics, Helmholtz Zentrum Muenchen, German Research Center for Environmental Health (GmbH), Ingolstaedter Landstr. 1, 85764, Neuherberg, Germany

    Frank Thiele, Christian M. Cohrs, Gerhard K. H. Przemeck, Helmut Fuchs & Martin Hrabé de Angelis

  2. Institute of Developmental Genetics, Helmholtz Zentrum Muenchen, German Research Center for Environmental Health (GmbH), Ingolstaedter Landstr. 1, 85764, Neuherberg, Germany

    Wolfgang Wurst

  3. Technische Universitaet Muenchen, Center of Life and Food Sciences Weihenstephan, Chair of Developmental Genetics, Alte Akademie 8, 85354, Freising, Germany

    Wolfgang Wurst

  4. Technische Universitaet Muenchen, Center of Life and Food Sciences Weihenstephan, Chair of Experimental Genetics, Alte Akademie 8, 85354, Freising, Germany

    Martin Hrabé de Angelis

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  1. Frank Thiele
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Correspondence to Martin Hrabé de Angelis.

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Thiele, F., Cohrs, C.M., Przemeck, G.K.H. et al. In vitro analysis of bone phenotypes in Col1a1 and Jagged1 mutant mice using a standardized osteoblast cell culture system. J Bone Miner Metab 31, 293–303 (2013). https://doi.org/10.1007/s00774-012-0421-x

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  • DOI: https://doi.org/10.1007/s00774-012-0421-x

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