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European Institute of Oncology

4 ARTICLES PUBLISHED IN JoVE

Biology

A Novel Method for the Culture and Polarized Stimulation of Human Intestinal Mucosa Explants

Katerina Tsilingiri¹, Angelica Sonzogni², Flavio Caprioli³, Maria Rescigno¹

¹Department of Experimental Oncology, European Institute of Oncology, ²Department of Pathology and Laboratory Medicine, European Institute of Oncology, ³U.O. Gastroenterologia 2, IRCCS Ca' Granda, Ospedale Policlinico di Milano

We introduce a novel method for the maintenance of human intestinal mucosa in culture and monitoring of the response to various types of stimuli over at least 24 hrs. With our method, the polarity of the tissue is maintained, allowing for a physiological stimulation via the apical route.

Biology

The ChroP Approach Combines ChIP and Mass Spectrometry to Dissect Locus-specific Proteomic Landscapes of Chromatin

Monica Soldi¹, Tiziana Bonaldi¹

¹Department of Experimental Oncology, European Institute of Oncology

By combining native and crosslinking chromatin immunoprecipitation with high-resolution Mass Spectrometry, ChroP approach enables to dissect the composite proteomic architecture of histone modifications, variants and non-histonic proteins synergizing at functionally distinct chromatin domains.

Immunology and Infection

Using Robotic Systems to Process and Embed Colonic Murine Samples for Histological Analyses

Fulvia Milena Cribiù^*1, Claudia Burrello^*2, Roberta Tacchi¹, Francesca Boggio¹, Dario Ricca¹, Flavio Caprioli^3,4, Stefano Ferrero¹, Federica Facciotti²

¹Pathology Unit, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, ²Department of Experimental Oncology, European Institute of Oncology, ³Gastroenterology and Endoscopy Unit, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, ⁴Department of Pathophysiology and Transplantation, Università degli Studi di Milano

Lack of standardization for murine tissue processing reduces the quality of murine histopathological analysis as compared to human specimens. Here, we present a protocol to perform histopathological examination of murine inflamed and uninflamed colonic tissues to show the feasibility of robotic systems routinely used for processing and embedding human samples.

Biochemistry

A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis

Marianna Maniaci^*1,2, Federica Marini^*1, Enrico Massignani^1,2, Tiziana Bonaldi¹

¹Department of Experimental Oncology, European Institute of Oncology IRCCS (IEO), ²European School of Molecular Medicine (SEMM), c/o Campus IFOM-IEO

Protein Arginine (R)-methylation is a wide-spread post-translational modification regulating multiple biological pathways. Mass spectrometry is the best technology to globally profile the R-methyl-proteome, when coupled to biochemical approaches for modified peptide enrichment. The workflow designed for the high confidence identification of global R-methylation in human cells is described here.