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Establishment and characterization of OS 99-1, a cell line derived from a highly aggressive primary human osteosarcoma

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Abstract

Osteosarcoma is the most common form of primary bone cancer. In this study, we established a human osteosarcoma cell line (OS 99-1) from a highly aggressive primary tumor. G-banding karyotype analysis demonstrated a large number of clonal abnormalities, as well as extensive intercellular heterogeneity. Through the use of immunologic, molecular, and biochemical analyses, we characterized protein and gene expression profiles confirming the osteogenic nature of the cells. Further evaluation indicated that OS 99-1 cells maintain the capacity to differentiate in an in vitro mineralization assay as well as form tumors in the in vivo chicken embryo model. This cell line provides a useful tool to investigate the molecular mechanisms contributing to osteosarcoma and may have the potential to serve as a culture system for studies involving bone physiology.

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Abbreviations

ALP:

tissue nonspecific alkaline phosphatase

ARS:

Alizarin Red S

BSP:

bone sialoprotein

LOH:

loss of heterogeneity

MTT:

3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide

OC:

osteocalcin

OS:

osteosarcoma

PVDF:

polyvinylidene fluoride

qRT-PCR:

quantitative real-time reverse transcriptase PCR

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Acknowledgments

We wish to thank Dr. Marileila Garcia (University of Colorado HSC) for her assistance with the karyotype analysis, Karen Helm (University of Colorado HSC) for her assistance with the flow cytometry, and Kate McInnerney and Dr. Jean Starkey (Montana State University) for their assistance and use of the equipment in the Genomics Core Facility. This work was supported in part by NIH R03 AR052898-01.

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Authors and Affiliations

  1. NICHD, Cell Biology and Metabolism Branch, National Institutes of Health, Bethesda, MD, 20892, USA

    Jennifer M. Gillette

  2. Department of Orthopaedics and Rehabilitation, University of Florida, Gainesville, FL, 32607, USA

    C. Parker Gibbs

  3. Department of Cell Biology and Neuroscience, Montana State University, PO Box 173148, Bozeman, MT, 59717, USA

    Sheila M. Nielsen-Preiss

Authors
  1. Jennifer M. Gillette
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  2. C. Parker Gibbs
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  3. Sheila M. Nielsen-Preiss
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Corresponding author

Correspondence to Sheila M. Nielsen-Preiss.

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Editor: J. Denry Sato

The work presented in this manuscript was initiated by the three authors at the University of Colorado Health Sciences Center, Denver, CO.

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Gillette, J.M., Gibbs, C.P. & Nielsen-Preiss, S.M. Establishment and characterization of OS 99-1, a cell line derived from a highly aggressive primary human osteosarcoma. In Vitro Cell.Dev.Biol.-Animal 44, 87–95 (2008). https://doi.org/10.1007/s11626-007-9075-8

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  • DOI: https://doi.org/10.1007/s11626-007-9075-8

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