Azienda Ospedaliera-Universitaria Careggi 2 articles published in JoVE Immunology and Infection Investigating Aortic Valve Calcification via Isolation and Culture of T Lymphocytes using Feeder Cells from Irradiated Buffy Coat Lavinia Curini*1,2, Mary Roxana Christopher*1, Herko Grubitzsch3,4, Ulf Landmesser1,3, Amedeo Amedei*2,5, Alexander Lauten*1,6, Brunilda Alushi*1,6 1Department of Cardiology, Campus Benjamin Franklin, Charité Universitätsmedizin Berlin and German Centre for Cardiovascular Research (DZHK), 2Department of Experimental and Clinical Medicine, University of Florence, 3Berlin Institute of Health, 4Department of Cardiology, German Heart Centre Berlin (DHZB), 5Sod of Interdisciplinary Internal Medicine, Azienda Ospedaliera Universitaria Careggi (AOUC), 6Department of General and Interventional Cardiology, Helios Klinikum Erfurt In this study, we describe the process of T lymphocyte isolation from fresh samples of calcified aortic valves and the analytical steps of T cell-cloning for the characterization of the adaptive leukocyte subsets by using flow cytometry analysis. Cancer Research Establishment of Cancer Stem Cell Cultures from Human Conventional Osteosarcoma Gaia Palmini1, Roberto Zonefrati1, Carmelo Mavilia1, Alessandra Aldinucci2, Ettore Luzi1, Francesca Marini1, Alessandro Franchi1, Rodolfo Capanna3, Annalisa Tanini1, Maria Luisa Brandi1 1Department of Surgery and Translational Medicine (DCMT), University of Florence, 2Neurofarba Department, University of Florence, 3Department of Traumatology and General Orthopedics, Azienda Ospedaliera Universitaria Careggi The presence of cancer stem cells (CSCs) in bone sarcomas has recently been linked to their pathogenesis. In this article, we present the isolation of CSCs from primary cell cultures obtained from human biopsies of conventional osteosarcoma (OS) using the ability of CSCs to grow under nonadherent conditions.