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Isolation and characterization of MSC from different sources (bone marrow, adipose tissue, decidual tissue derived MSC from endometrial decidual tissue)
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Isolation and characterization of human T lymphocytes (CTL)
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Isolation and characterization of tumor stromal cells
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Ex vivo amplification, introducing both standard and innovative culture conditions: -different media and proprietary media both for expansion and differentiation; -use of different culture supports (plates, flasks, bags, matrices); development of 3D tissue like-cultures from either cell lines and primary cells by VITVO platform-Rigenerand, lymphocytes expansion through bioreactors (Wave Bioreactor, System 2/10 EH, GE-Healtcare Life Science)
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Immunophenotyping and sorting by FACS (FACSAria III, Becton Dickinson) or MACS Cell Separation (Miltenyi Biotec)
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Isolation and characterization by FACS and ZetaView technology of Micro, Nano Vesicles and Exosomes from cell lines and primary cells from patients, including either cancer and stem cells
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Time-lapse microscopy (Zeiss Axiovert 200M)
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Proliferation and migration assays (Cell Proliferation Assay by FACS analysis & BrdU, CFSE or propidium iodide staining; 96-well plate proliferation assays: MTS assay, fluorescent assay, luminescent cell viability assay; transwell systems, time lapse microscopy)
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Potency assays (cytochemical and molecular assay)
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Cytotoxicity assays both in vitro (FACS analyses: detection and quantification of apoptotic/necrotic cells by Supervital Propidium Iodine, Annexin V, activated Caspase3-8 staining; 51Cr release by Microbeta 2 Perkin Elmer; Cell Viability Assay on the GloMax® Discover System-Pormega; time lapse microscopy) and in vivo in NOD-SCID mice
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Plasmid vector construction for cell gene-modification and protein expression (FACS analyses)
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Gene manipulation of mesenchymal stromal/stem cells and lymphocytes by viral vectors (CAR-T approach)
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Western Blotting and in-cell Western by LI-COR Odissey Infrared Imaging System
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Orthotopic (i.e. pancreatic and glioma tumors), Metastatic (Ewing’s Sarcoma) and Heterotopic, Xenotraplant models in NOD-SCID and NSG mice
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Pre-clinical in vivo models of tissue regeneration combining cells and biomaterials both in immunocompetent and NOD-scid mice
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Pre-clinical in vivo testing of biocompatibility and vascularization
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Multiparametric histological analyses and microscopy
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Early clinical implementation and regulatory requirement
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Safety profile analyses
