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JoVE Journal Cancer Research
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Obtaining Cancer Stem Cell Spheres from Gynecological and Breast Cancer Tumors

Article DOI: 10.3791/60022-v • 07:01 min • March 1st, 2020
March 1st, 2020 •
*1,2,3,4, *1,2,3,4,5,6, 1,2,3, 2,7, 1,2,3,4,8, 9, 3,9,10, 1,2,3,4,8
1Institute of Biophysics, Faculty of Medicine, University of Coimbra, 2Coimbra Institute for Clinical and Biomedical Research (iCBR), area of Environment Genetics and Oncobiology (CIMAGO), Faculty of Medicine, University of Coimbra, 3CNC.IBILI/Center for Innovative Biomedicine and Biotechnology (CIBB), University of Coimbra, 4Clinical Academic Center of Coimbra (CACC), 5Universitary Clinic of Gynecology, Faculty of Medicine, University of Coimbra, 6Gynecology A Service, Coimbra Hospital and Universitary Center, 7Institute of Pharmacology & Experimental Therapeutics, Faculty of Medicine, University of Coimbra, 8Institute of Experimental Pathology, Faculty of Medicine, University of Coimbra, 9Cytometry Operational Management Unit, Clinical Pathology Service, Coimbra Hospital and Universitary Center, 10Polytechnic Institute of Coimbra, ESTESC-Coimbra Health School, Laboratory Biomedical Sciences
* These authors contributed equally

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Summary
Automatic Translation

The aim of this methodology is to identify cancer stem cells (CSC) in cancer cell lines and primary human tumor samples with the sphere-forming protocol, in a robust manner, using functional assays and phenotypic characterization with flow cytometry and Western blot.

Tags

Cancer Stem Cells Gynecological Cancer Breast Cancer Tumor Isolation Functional Assays Phenotypic Characterization Therapies Resistance Reoccurrence Non-adherent Suspension Cultures PolyHEMA Coating Trypsin EDTA Cell Culture Medium Centrifugation Sphere Culturing Medium Methyl Cellulose Anchorage-free Environment Incubation
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