Centro Cardiologico Monzino IRCCS

3 ARTICLES PUBLISHED IN JoVE

Biochemistry

Optimized Protocol for the Extraction of Proteins from the Human Mitral Valve

Cristina Banfi¹, Anna Guarino², Maura Brioschi¹, Stefania Ghilardi¹, Valeria Mastrullo¹, Elena Tremoli¹, 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.

Biology

Microvascular and Macrovascular Endothelial Cell Isolation and Purification from Lung-Derived Samples

Roberto Plebani^1,2, Alessandra D'Alessandro², Paola Lanuti^2,3, Pasquale Simeone^2,3, Massimiliano Cinalli¹, Ilaria Righi⁴, Alessandro Palleschi^4,5, Matteo Mucci^1,2, Marco Marchisio^2,3, Francesco Cappabianca¹, Marina Camera^6,7, Felice Mucilli¹, 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.

Biology

Flow Cytometry Analysis of Tissue Factor Expression in Human Platelets

Marta Brambilla¹, Alessia Becchetti¹, Kevin Nallio¹, 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.