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Complutense University

5 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Phenotypic Characterization of Macrophages from Rat Kidney by Flow Cytometry
Alfonso Rubio-Navarro 1, Melania Guerrero-Hue 1, Beatriz Martín-Fernandez 2, Isabel Cortegano 3, Elena Olivares-Alvaro 2, Natalia de las Heras 2, Mario Alía 3, Belén de Andrés 3, María Luisa Gaspar 3, Jesús Egido 1, Juan Antonio Moreno 1
1Renal, Vascular and Diabetes Research Lab, IIS-Fundaciòn Jiménez Dìaz, Autonoma University, 2Department of Physiology, Faculty of Medicine, Complutense University, 3Department of Immunology, Centro Nacional de Microbiologìa, Instituto de Salud Carlos III (ISCIII)

This manuscript describes a detailed protocol for phenotypic and quantitative analysis of resident macrophages from rat kidneys by flow cytometry. The resulting stained cells can be also used for other applications, including cell sorting, gene expression analysis or functional studies, thus increasing the information obtained in the experimental model.

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Neuroscience

Live Imaging Followed by Single Cell Tracking to Monitor Cell Biology and the Lineage Progression of Multiple Neural Populations
Rosa Gómez-Villafuertes *1,2,3, Lucía Paniagua-Herranz *1,2,3, Sergio Gascon *4,5, David de Agustín-Durán 1,2,3, María de la O Ferreras 1,2,3, Juan Carlos Gil-Redondo 1,2,3, María José Queipo 1,2,3, Aida Menendez-Mendez 1,2,3, Ráquel Pérez-Sen 1,2,3, Esmerilda G. Delicado 1,2,3, Javier Gualix 1,2,3, Marcos R. Costa 6, Timm Schroeder 7, María Teresa Miras-Portugal 1,2,3, Felipe Ortega 1,2,3
1Biochemistry and Molecular Biology Department, Faculty of Veterinary medicine, Complutense University, 2University Institute for Neurochemistry Research (IUIN), 3Instituto de Investigación Sanitaria del Hospital Clínico San Carlos (IdISSC), 4Institute of Stem Cell Research, Helmholtz Center Munich, Neuherberg/Munich, Germany Physiological Genomics, Biomedical Center, Ludwig-Maximilians University Munich, 5Toxicology and Pharmacology Department, Faculty of Veterinary medicine, Complutense University, 6Brain Institute, Federal University of Rio Grande do Norte, 7Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule (ETH) Zurich

A robust protocol to monitor neural populations by time-lapse video-microscopy followed by software-based post-processing is described. This method represents a powerful tool to identify biological events in a selected population during live imaging experiments.

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Biology

Stimulation of Stem Cell Niches and Tissue Regeneration in Mouse Skin by Switchable Protoporphyrin IX-Dependent Photogeneration of Reactive Oxygen Species In Situ
Jesús Espada 1,2, Elisa Carrasco 3, María I. Calvo-Sánchez 1,4, Sandra Fernández-Martos 1, Juan José Montoya 5
1Ramón y Cajal Institute for Health Research (IRYCIS), Ramón y Cajal University Hospital, 2Centro Integrativo de Biología y Química Aplicada (CIBQA), Universidad Bernardo O'Higgins, 3Department of Molecular Biology, Faculty of Sciences, Universidad Autónoma de Madrid (UAM), 4Instituto de Investigaciones Biosanitarias, Facultad de Ciencias Experimentales, Universidad Francisco de Vitoria, 5Department of Radiology, Rehabilitation & Physiotherapy, Faculty of Medicine, Complutense University

The aim of this protocol is to induce transient in vivo production of nonlethal levels of reactive oxygen species (ROS) in mouse skin, further promoting physiological responses in the tissue.

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Neuroscience

Double In Utero Electroporation to Target Temporally and Spatially Separated Cell Populations
Isabel Mateos-White 1, Jaime Fabra-Beser 1, David de Agustín-Durán 1, Cristina Gil-Sanz 1
1Estructura de Recerca Interdisciplinar en Biotecnología y Biomedicina (ERI BIOTECMED), Departamento de Biología Celular, Biología Funcional y Antropología Física, Universidad de Valencia

Double in utero electroporation allows targeting cell populations that are spatially and temporally separated. This technique is useful to visualize interactions between those cell populations using fluorescent proteins in normal conditions but also after functional experiments to perturb genes of interest.

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Bioengineering

Quantitative MRI of Endothelial Permeability and (Dys)function in Atherosclerosis
Begoña Lavin 1,2,3, Marcelo E. Andia 1,4,5, Prakash Saha 6, René M. Botnar 1,2,7,8, Alkystis Phinikaridou 1,2
1School of Biomedical Engineering and Imaging Sciences, King’s College London, 2BHF Centre of Excellence, Cardiovascular Division, King’s College London, 3Biochemistry and Molecular Biology Department, School of Chemistry, Complutense University, 4Radiology Department, School of Medicine, Pontificia Universidad Católica de Chile, 5ANID - Millennium Science Initiative Program - Millennium Nucleus for Cardiovascular Magnetic Resonance, 6Academic Department of Vascular Surgery, Cardiovascular Division, King’s College London, 7Wellcome Trust and EPSRC Medical Engineering Center, King’s College London, 8Escuela de Ingeniería, Universidad Católica de Chile

We have developed an accurate, non-invasive, and easy-to-use method to quantify endothelial permeability and dysfunction in the arteries using Magnetic Resonance Imaging (MRI), named qMETRIC. This technique enables assessing vascular damage and cardiovascular risk associated with atherosclerosis in preclinical models and humans.

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