IRCCS 3 articles published in JoVE Cancer Research Live-3D-Cell Immunocytochemistry Assays of Pediatric Diffuse Midline Glioma Giulia Pericoli*1, Roberta Ferretti*1, Andrew S. Moore2,3, Maria Vinci1 1Department of Onco-hematology, Gene and Cell Therapy, Bambino Gesù Children's Hospital, IRCCS, 2 This study presents a protocol of live-3D-cell immunocytochemistry applied to a pediatric diffuse midline glioma cell line, useful to study in real-time the expression of proteins on the plasma membrane during dynamic processes like 3D cell invasion and migration. Immunology and Infection A DNA/Ki67-Based Flow Cytometry Assay for Cell Cycle Analysis of Antigen-Specific CD8 T Cells in Vaccinated Mice Sonia Simonetti*1,2, Ambra Natalini*1,2, Giovanna Peruzzi3, Alfredo Nicosia4, Antonella Folgori5, Stefania Capone5, Angela Santoni2,6, Francesca Di Rosa1 1Institute of Molecular Biology and Pathology, National Research Council of Italy (CNR), 2Department of Molecular Medicine, University of Rome “Sapienza”, 3Center for Life Nano Science, Istituto Italiano di Tecnologia, 4Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 5Reithera Srl, 6IRCCS, Neuromed Clonal expansion is a key feature of antigen-specific T cell response. However, the cell cycle of antigen-responding T cells has been poorly investigated, partly because of technical limitations. We describe a flow cytometric method to analyze clonally expanding antigen-specific CD8 T cells in spleen and lymph nodes of vaccinated mice. Cancer Research Development of a Human Preclinical Model of Osteoclastogenesis from Peripheral Blood Monocytes Co-cultured with Breast Cancer Cell Lines Laura Mercatali1, Chiara Spadazzi1, Giacomo Miserocchi1, Chiara Liverani1, Alessandro De Vita1, Alberto Bongiovanni1, Federica Recine1, Dino Amadori1, Toni Ibrahim1 1Osteoncology and Rare Tumors Center, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST), IRCCS This protocol describes development of an in vitro human preclinical model of osteoclastogenesis from peripheral blood monocytes cultured with breast cancer cell lines to mimic the cancer cell-osteoclast interaction. The model could be used to further our understanding of bone metastasis formation and improve therapeutic options.
