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Centro Cardiologico Monzino IRCCS

3 ARTICLES PUBLISHED IN JoVE

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Biochemistry

Optimized Protocol for the Extraction of Proteins from the Human Mitral Valve
Cristina Banfi 1, Anna Guarino 2, Maura Brioschi 1, Stefania Ghilardi 1, Valeria Mastrullo 1, Elena Tremoli 1, Gianluca Polvani 2,3,4
1Centro Cardiologico Monzino IRCCS, 2Cardiovascular Tissue Bank of Milan, Centro Cardiologico Monzino IRCCS, 3Department of Clinical Sciences and Community Health, Cardiovascular Section, University of Milan, 4Department of Cardiovascular Disease, Development and Innovation Cardiac Surgery Unit, Centro Cardiologico Monzino IRCCS

The protein composition of the human mitral valve is still partially unknown, because its analysis is complicated by low cellularity and therefore by low protein biosynthesis. This work provides a protocol to efficiently extract protein for the analysis of the mitral valve proteome.

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Biology

Microvascular and Macrovascular Endothelial Cell Isolation and Purification from Lung-Derived Samples
Roberto Plebani 1,2, Alessandra D'Alessandro 2, Paola Lanuti 2,3, Pasquale Simeone 2,3, Massimiliano Cinalli 1, Ilaria Righi 4, Alessandro Palleschi 4,5, Matteo Mucci 1,2, Marco Marchisio 2,3, Francesco Cappabianca 1, Marina Camera 6,7, Felice Mucilli 1, Mario Romano 1,2
1Department of Medical, Oral and Biotechnological Sciences, “G. d’Annunzio” University of Chieti-Pescara, 2Center on Advanced Studies and Technology (CAST), “G. d’Annunzio” University of Chieti-Pescara, 3Department of Medicine and Aging Sciences, University “G. d’Annunzio” of Chieti-Pescara, 4U.O. di Chirurgia Toracica e dei Trapianti di Polmone, Fondazione IRCCS Ca’ Granda - Ospedale Maggiore Policlinico di Milano, 5Dipartimento di Fisiopatologia Medico-Chirurgica e dei Trapianti, Università degli Studi di Milano, 6Department of Pharmaceutical Sciences, Università degli Studi di Milano, 7Unit of Cell and Molecular Biology in Cardiovascular Diseases, Centro Cardiologico Monzino IRCCS

Although challenging, the isolation of pulmonary endothelial cells is essential for studies on lung inflammation. The present protocol describes a procedure for the high-yield, high-purity isolation of macrovascular and microvascular endothelial cells.

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Biology

Flow Cytometry Analysis of Tissue Factor Expression in Human Platelets
Marta Brambilla 1, Alessia Becchetti 1, Kevin Nallio 1, Marina Camera 1,2
1Centro Cardiologico Monzino IRCCS, 2Department of Pharmaceutical Sciences, Università degli Studi di Milano

This article outlines the protocol for quantifying the percentage of Tissue Factor (TF)-positive platelets using whole blood flow cytometry, assessing the protein: (1) intracellularly in resting conditions, and (2) on the cell surface, in both resting and activated conditions. Guidance is also provided for evaluating TF-positive platelets in platelet-rich plasma.

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