Establishment of Cancer Stem Cell Cultures from Human Conventional Osteosarcoma

被引:9
作者
Palmini, Gaia [1 ]
Zonefrati, Roberto [1 ]
Mavilia, Carmelo [1 ]
Aldinucci, Alessandra [2 ]
Luzi, Ettore [1 ]
Marini, Francesca [1 ]
Franchi, Alessandro [1 ]
Capanna, Rodolfo [3 ]
Tanini, Annalisa [1 ]
Brandi, Maria Luisa [1 ]
机构
[1] Univ Florence, Dept Surg & Translat Med DCMT, I-50121 Florence, Italy
[2] Univ Florence, Neurofarba Dept, I-50121 Florence, Italy
[3] Azienda Ospedaliera Univ Careggi, Dept Traumatol & Gen Orthoped, Florence, Italy
来源
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS | 2016年 / 116期
关键词
Cancer Research; Issue; 116; Cellular biology; cancer; bone sarcomas; conventional osteosarcoma; cancer stem cells; sphere formation assay; 3-dimensional cell culture; sarcospheres; osteosarcoma cancer stem cells; stem cell markers; mesenchymal stem cells; embryonic stem cells; ACUTE MYELOID-LEUKEMIA; OSTEOGENIC-SARCOMA; DRUG-RESISTANCE; SOLID TUMORS; DIFFERENTIATION; CHILDHOOD; BONE; DEDIFFERENTIATION; TRANSFORMATION; HETEROGENEITY;
D O I
10.3791/53884
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
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页数:17
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