Abstract 19

A NEW GFP EXPRESSING OSTEOSARCOMA ANIMAL MODEL

Joanna Lenaghan, Karla Contreras, Anne Nadesapillai, Peter Choong, Joseline Ojaimi

Department of Orthopaedics, The University of Melbourne, St. Vincent's Hospital, Melbourne

25-40% of patients suffering from osteosarcoma will succumb to the disease, an overwhelming 98% from pulmonary metastases. Better understanding of micrometastasis holds the key to improved survival outcomes. Our aim has been to develop an animal model of osteosarcoma that will be amenable to more sophisticated imaging, providing 'illumination' of micrometastases and their pathogenesis.

U2OS, a non-transformed, human osteosarcoma cell line was stably transfected with green fluorescent protein (GFP). Cells were selected in G418 for 4 weeks to enrich GFP expression. Transfected and untransfected cell lines were analysed in vitro to determine the effects of transfection on the cells proliferative and migratory properties. Balb-c nude mice were inoculated with the U2OS-eGFP cell line using intratibial injection, and were sacrificed at 2, 4 and 6 weeks. Post inoculation the tibia and lungs were examined intact, and as histological sections, under fluorescent microscope.

The model has been successful in establishment of GFP expressing tumour, identifiable under fluorescent light. This important animal model is unique in the study of osteosarcoma. It will enable more accurate and easier analysis of tumour evolution in vivo, particularly in the detection and study of micrometastasis.

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