University of Porto

5 articles published in JoVE

• Cancer Research

  
  Yeast As a Chassis for Developing Functional Assays to Study Human P53 Paola Monti¹, Bartolomeo Bosco², Sara Gomes³, Lucilia Saraiva*³, Gilberto Fronza*¹, Alberto Inga*^{2 1}Mutagenesis and Cancer Prevention Unit, Ospedale Policlinico San Martino, ²Department CIBIO, University of Trento, ³LAQV/REQUIMTE, Laboratory of Microbiology, Faculty of Pharmacy, University of Porto Presented here are four protocols to construct and exploit yeast Saccharomyces cerevisiae reporter strains to study human P53 transactivation potential, impacts of its various cancer-associated mutations, co-expressed interacting proteins, and the effects of specific small molecules.

• Bioengineering

  
  Comparable Decellularization of Fetal and Adult Cardiac Tissue Explants as 3D-like Platforms for In Vitro Studies Ana C. Silva^1,2,3,4, Maria J. Oliveira^1,2,5, Todd C McDevitt^4,6, Mário A. Barbosa^1,2,3, Diana S. Nascimento^1,2, Perpétua Pinto-do-Ó^{1,2,3 1}i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, ²INEB - Instituto de Engenharia Biomédica, Universidade do Porto, ³Instituto de Ciências Biomédicas Abel Salazar (ICBAS), Universidade do Porto, ⁴Gladstone Institute of Cardiovascular Disease, ⁵Faculty of Medicine, University of Porto, ⁶University of California San Francisco The cardiac extracellular matrix (ECM) is a complex network of molecules that orchestrate key processes in tissues and organs while enduring physiological remodeling throughout life. Standardized decellularization of fetal and adult hearts permits comparative experimental studies of both tissues in a 3D context by capturing native architecture and biomechanical properties.

• Neuroscience

  
  Modified Roller Tube Method for Precisely Localized and Repetitive Intermittent Imaging During Long-term Culture of Brain Slices in an Enclosed System Benjamin B. Fixman*¹, Isaac W. Babcock*¹, Laurie S. Minamide*¹, Alisa E. Shaw¹, Marina I. Oliveira da Silva^1,2, Avery M. Runyan¹, Michael T. Maloney^1,3, Jeffrey J. Field¹, James R. Bamburg^{1 1}Department of Biochemistry and Molecular Biology and Molecular, Cellular and Integrated Neuroscience Program, Colorado State University, ²IBMC-Instituto de Biologia Molecular e Celular, i3S-Instituto de Investigaçãoe Inovação em Saúde, ICBAS, Universidade do Porto, ³Denali Therapeutics Presented here is a modified roller tube method for culturing and intermittent high-resolution imaging of rodent brain slices over many weeks with precise repositioning on photoetched coverslips. Neuronal viability and slice morphology are well maintained. Applications of this fully enclosed system using viruses for cell-type specific expression are provided.

• Developmental Biology

  
  A Standardized Approach for Multispecies Purification of Mammalian Male Germ Cells by Mechanical Tissue Dissociation and Flow Cytometry Ana C. Lima*^1,2,3,4, Min Jung*¹, Jannette Rusch¹, Abul Usmani¹, Alexandra M. Lopes^3,4, Donald F. Conrad^{1 1}Department of Genetics, Washington University School of Medicine, ²Graduate Program in Areas of Basic and Applied Biology (GABBA), Abel Salazar Institute of Biomedical Sciences, University of Porto, ³Instituto de Investigação e Inovação em Saúde, University of Porto, ⁴IPATIMUP - Instituto de Patologia e Imunologia Molecular da Universidade do Porto, ⁵Department of Pathology & Immunology, Washington University School of Medicine This work describes the standardization of a method to obtain purified germ cell populations from testicular tissue of different mammalian species. It is a straightforward protocol that combines mechanical testis dissociation, staining with Hoechst-33342 and propidium iodide, and FACS sorting, with wide applications in comparative studies of male reproductive biology.

• Biology

  
  Analysis of Cell Suspensions Isolated from Solid Tissues by Spectral Flow Cytometry Sandrine Schmutz¹, Mariana Valente^2,3,4,5, Ana Cumano², Sophie Novault^{1 1}Flow Cytometry Core Facility, Center for Translational Research-Technical Core, Institut Pasteur, ²Unit for Lymphopoiesis, Immunology Department, INSERM U1223, University Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Institut Pasteur, ³Stem-Cell Microenvironments in Repair/Regeneration Team, Instituto de Investigação e Inovação em Saúde (i3s), INEB - Instituto de Engenharia Biomédica, ⁴ICBAS - Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, ⁵Stem Cells and Regenerative Medicine Team, UMRS 1166, ICAN - Institute of Cardiometabolism And Nutrition, UPMC - Université Pierre et Marie Curie - Paris 6, INSERM This article describes spectral cytometry, a new approach in flow cytometry that uses the shapes of emission spectra to distinguish fluorochromes. An algorithm replaces compensations and can treat auto-fluorescence as an independent parameter. This new approach allows for the proper analysis of cells isolated from solid organs.