Humanitas Clinical and Research Center 4 articles published in JoVE Cancer Research Isolation of Proximal Fluids to Investigate the Tumor Microenvironment of Pancreatic Adenocarcinoma Greta Donisi*1, Marialuisa Barbagallo*2, Giovanni Capretti1,3, Gennaro Nappo1,3, Panteleimon G. Takis4,5,6, Alessandro Zerbi1,3, Federica Marchesi2,7, Nina Cortese2 1Section of Pancreatic Surgery, Humanitas Clinical and Research Center-IRCCS, 2Department of Immunology and Inflammation, Humanitas Clinical and Research Center-IRCCS, 3Department of Biomedical Sciences, Humanitas University, 4Giotto Biotech S.R.L., 5Section of Bioanalytical Chemistry, Division of Systems Medicine, Department of Metabolism, Digestion and Reproduction, Imperial College London, 6National Phenome Centre, Department of Metabolism, Digestion and Reproduction, Faculty of Medicine, Imperial College London, 7Department of Medical Biotechnology and Translational Medicine, University of Milan Pancreatic juice is a precious source of biomarkers for human pancreatic cancer. We describe here a method for intraoperative collection procedure. To overcome the challenge of adopting this procedure in murine models, we suggest an alternative sample, tumor interstitial fluid, and describe here two protocols for its isolation. Biology Interactome-Seq: A Protocol for Domainome Library Construction, Validation and Selection by Phage Display and Next Generation Sequencing Maria Felicia Soluri1, Simone Puccio2, Giada Caredda2, Giorgio Grillo3, Vito Flavio Licciulli3, Arianna Consiglio3, Paolo Edomi4, Claudio Santoro1, Daniele Sblattero4, Clelia Peano5,6 1Department of Health Sciences, Università del Piemonte Orientale & IRCAD, Novara, Italy, 2Institute of Biomedical Technologies, National Research Council, Segrate, Milan, Italy, 3Institute of Biomedical Technologies, National Research Council, Bari, Italy, 4Department of Life Sciences, University of Trieste, Italy, 5Institute of Genetic and Biomedical Research, National Research Council, Rozzano, Milan, Italy, 6Humanitas Clinical and Research Center, Rozzano, Milan, Italy The protocols described allow the construction, characterization and selection (against the target of choice) of a "domainome" library made from any DNA source. This is achieved by a research pipeline that combines different technologies: phage display, a folding reporter and next generation sequencing with a web tool for data analysis. Immunology and Infection Preparation of Single-cell Suspensions for Cytofluorimetric Analysis from Different Mouse Skin Regions Achille Broggi1,2, Clara Cigni1, Ivan Zanoni1,2,3, Francesca Granucci1,3 1Department of Biotechnology and Biosciences, University of Milano-Bicocca, 2Boston Children's Hospital, Division of Gastroenterology, Harvard Medical School, 3Humanitas Clinical and Research Center The skin is home to a complex immune cell network. We describe an efficient methodology for the digestion of mouse skin, from different parts of the animal's body, in order to obtain a single-cell suspension and analyze the different leukocyte populations resident in the skin by flow cytometry. Biology Generation of Human Cardiomyocytes: A Differentiation Protocol from Feeder-free Human Induced Pluripotent Stem Cells Elisa Di Pasquale1,2, Belle Song1, Gianluigi Condorelli1 1Humanitas Clinical and Research Center, Italy, 2Institute of Genetic and Biomedical Research (IRGB), National Research Council (CNR) Pluripotent stem cells, either embryonic or induced pluripotent stem (iPS) cells, constitute a valuable source of human differentiated cells, including cardiomyocytes. Here, we will focus on cardiac induction of iPS cells, showing how to use them to obtain functional human cardiomyocytes through an embryoid bodies-based protocol.
