title: Establishment of Cancer Stem Cell Cultures from Human Conventional Osteosarcoma.
abstract: The current improvements in therapy against osteosarcoma (OS) have prolonged the lives of cancer patients, but the survival rate of five years remains poor when metastasis has occurred. The Cancer Stem Cell (CSC) theory holds that there is a subset of tumor cells within the tumor that have stem-like characteristics, including the capacity to maintain the tumor and to resist multidrug chemotherapy. Therefore, a better understanding of OS biology and pathogenesis is needed in order to advance the development of targeted therapies to eradicate this particular subset and to reduce morbidity and mortality among patients. Isolating CSCs, establishing cell cultures of CSCs, and studying their biology are important steps to improving our understanding of OS biology and pathogenesis. The establishment of human-derived OS-CSCs from biopsies of OS has been made possible using several methods, including the capacity to create 3-dimensional stem cell cultures under nonadherent conditions. Under these conditions, CSCs are able to create spherical floating colonies formed by daughter stem cells; these colonies are termed "cellular spheres". Here, we describe a method to establish CSC cultures from primary cell cultures of conventional OS obtained from OS biopsies. We clearly describe the several passages required to isolate and characterize CSCs.
authors: Department of Surgery and Translational Medicine (DCMT), University of Florence.
authors: Gaia
authors: G
authors: Palmini
authors: Department of Surgery and Translational Medicine (DCMT), University of Florence.
authors: Roberto
authors: R
authors: Zonefrati
authors: Department of Surgery and Translational Medicine (DCMT), University of Florence.
authors: Carmelo
authors: C
authors: Mavilia
authors: Neurofarba Department, University of Florence.
authors: Alessandra
authors: A
authors: Aldinucci
authors: Department of Surgery and Translational Medicine (DCMT), University of Florence.
authors: Ettore
authors: E
authors: Luzi
authors: Department of Surgery and Translational Medicine (DCMT), University of Florence.
authors: Francesca
authors: F
authors: Marini
authors: Department of Surgery and Translational Medicine (DCMT), University of Florence.
authors: Alessandro
authors: A
authors: Franchi
authors: Department of Traumatology and General Orthopedics, Azienda Ospedaliera Universitaria Careggi.
authors: Rodolfo
authors: R
authors: Capanna
authors: Department of Surgery and Translational Medicine (DCMT), University of Florence.
authors: Annalisa
authors: A
authors: Tanini
authors: Department of Surgery and Translational Medicine (DCMT), University of Florence; marialuisa.brandi@unifi.it.
authors: Maria Luisa
authors: ML
authors: Brandi
mesh_terms: D001859:Bone Neoplasms; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D006801:Humans; D014411:Neoplastic Stem Cells; D012516:Osteosarcoma