ICCBH2017 Poster Presentations (1) (209 abstracts)
Development of an in vitro model of cancer stem cells from a rare human telangiectatic osteosarcoma
Gaia Palmini 1 , Roberto Zonefrati 1 , Cecilia Romagnoli 1 , Alessandra Aldinucci 2 , Gianna Galli 1 , Carmelo Mavilia 1 , Gigliola Leoncini 1 , Antonella Simoni 1 , Alessandro Franchi 1 , Domenico Andrea Campanacci 3 , Rodolfo Capanna 4 & Maria Luisa Brandi 1
1Department of Surgery and Translational Medicine, University of Florence, Florence, Italy; 2Department of Neurofarba, University of Florence, Florence, Italy; 3SOD Ortopedia Oncologica e Ricostruttiva, AOU Careggi, Florence, Italy; 4Clinica universitaria Ortopedia e Traumatologia, Azienda Ospedaliera, Pisa, Italy.
Objective: Even though recent studies have proved the presence of cancer stem cells (CSCs) in osteosarcoma (OSA), with this study, for the first time, the existence of CSCs in a rare high grade type of OSA, the telangiectatic osteogenic sarcoma (TOS), is showed.
Methods: TOS sample was collected at the ‘Ortopedia Oncologica e Ricostruttiva Unit’, AOU Careggi, Florence, with informed consent approved by the local Ethical Committee. First of all, the primary human cancer cell culture of TOS has been established. After that, the subpopulation of CSCs has been isolated from this by the sarcosphere formation assay. Consequently, several cellular assays/stainings and molecular analyses have been performed to assess the presence of markers and properties, which are unique signature of the cancer stem cells phenotype.
Results: We have set up a primary cell line of a high grade TOS, from which we have isolated the CSCs and we have obtained a TOS-CSCs line, called TOS1-CSCs. The cancer stem cells phenotype of TOS1-CSC line was confirmed by observing the capacity of the TOS1-CSCs to differentiate into osteoblasts and into adipocytes and by showing the positive presence of the mesenchymal stem cells (MSCs) markers (by immunofluorescence assays and by the flow cytometry, too. The TOS1-CSCs line has showed a good rate as clonogenic capacity and a good rate as tumorigenic capacity, which has been evaluated in vitro by the agar soft assay. We have also studied, and confirmed, their embryonic phenotype by verifying the presence of the expression of the embryonic stem cells (ESCs) marker genes together with CD133 other genes involved in the pluripotency of CSCs and in the metastatic process.
Conclusions: In conclusion we have established and completely characterized, for the first time, a TOS-CSCs line at cellular and molecular level, setting up an in vitro model to study/find new targets to permit the development of molecular therapy against this high grade type of OSA.
Acknowledgements: This research has been supported by the Istituto Toscano Tumori (ITT) _Regione Toscana_Grant Proposal 2010.
Disclosure: The authors declared no competing interests.
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