Tumorsphere Derivation and Treatment from Primary Tumor Cells Isolated from Mouse Rhabdomyosarcomas
| Autor: | Alessandra Sacco, Francesca Boscolo Sesillo |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Male
Candidate gene genetic structures Carcinogenesis General Chemical Engineering Cell Culture Techniques Mice SCID 030204 cardiovascular system & hematology Biology Muscle Development medicine.disease_cause Article General Biochemistry Genetics and Molecular Biology Mice 03 medical and health sciences 0302 clinical medicine Mice Inbred NOD In vivo Rhabdomyosarcoma 0502 economics and business Cell Adhesion Tumor Cells Cultured medicine Animals Humans General Immunology and Microbiology General Neuroscience Soft tissue sarcoma 05 social sciences Sarcoma Transfection medicine.disease Primary tumor Recombinant Proteins Cell culture Mutation Cancer research Female 050211 marketing |
| Zdroj: | J Vis Exp |
| ISSN: | 1940-087X |
| Popis: | Rhabdomyosarcoma is the most common soft tissue sarcoma in children. Although significant efforts enabled the identification of common mutations associated with RMS, and allowed discrimination of different RMS subtypes, major challenges still exist for the development of novel treatments to further improve patients’ prognosis. Although identified by the expression of myogenic markers, there is still significant controversy on whether RMS has myogenic or non-myogenic origin, as the cell of origin is still poorly understood. In the present study, we provide a reliable method for the tumorsphere assay for mouse RMS. The assay is based on functional properties of tumor cells and allows the identification of rare populations within the tumor with tumorigenic function. We also describe procedures for testing recombinant proteins and integrating transfection protocols with the tumorsphere assay, for evaluating candidate genes involved in tumor development and growth. We further describe a procedure for allograft transplantation of tumorspheres into recipient mice, to validate tumorigenic function in vivo. Overall, the described method allows reliable identification and testing of rare RMS tumorigenic populations that can be applied to RMS arising in different contexts. Finally, the protocol can be utilized as a platform for drug screening and future development of therapeutics. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|