3 articles published in JoVE
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 Polvani2,3,4 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.
Analysis of Chromosome Segregation, Histone Acetylation, and Spindle Morphology in Horse Oocytes Federica Franciosi1, Irene Tessaro1,2, Rozenn Dalbies-Tran3, Cecile Douet3, Fabrice Reigner4, Stefan Deleuze5, Pascal Papillier3, Ileana Miclea6, Valentina Lodde1, Alberto M. Luciano1, Ghylene Goudet3 1Department of Health, Animal Science and Food Safety, University of Milan, 2IRCCS. Istituto Ortopedico Galeazzi, 3PRC, CNRS, IFCE, Université de Tours, INRA, 4PAO, INRA, 5Clinique des Animaux de Compagnie et des Équidés, Université de Liège, 6University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca, Romania This manuscript describes an experimental approach to morphologically and biochemically characterize horse oocytes. Specifically, the present work illustrates how to collect immature and mature horse oocytes by ultrasound-guided ovum pick-up (OPU) and how to investigate chromosome segregation, spindle morphology, global histone acetylation, and mRNA expression.
Phenotypic Analysis and Isolation of Murine Hematopoietic Stem Cells and Lineage-committed Progenitors Michela Frascoli1, Michele Proietti1, Fabio Grassi1,2 1Institute for Research in Biomedicine, Bellinzona (Switzerland), 2Dipartimento di Biologia e Genetica per le Scienze Mediche, Universitá degli Studi di Milano A method to analyse the distribution of bone marrow hematopoietic progenitors in flow cytometry as well as to efficiently isolate highly purified hematopoietic stem cells (HSCs) is described. The isolation procedure is essentially based on magnetic enrichment of c-Kit+ cells and cell sorting to purify HSCs for cellular and molecular studies.