Maastricht University

5 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells

Gisela Nogales-Gadea^*1,2, Abhishek Saxena^*1, Carolin Hoffmann^*1, Judith Hounjet¹, Daniëlle Coenen¹, Peter Molenaar¹, Mario Losen¹, Pilar Martinez-Martinez¹

¹School for Mental Health and Neuroscience, Maastricht University, ²Department of Neurosciences, Institut d'Investigació Germans Trias i Pujol

Synthesis of human monoclonal antibodies is the first step in studies aimed at unraveling the pathophysiological mechanisms of auto-antibody-mediated immune responses. We have developed a protocol to generate recombinant human immunoglobulin G (IgG) monoclonal antibodies from blood sorted B cells, including B-cell isolation, antibody cloning and in vitro synthesis.

Immunology and Infection

Laminar Flow-based Assays to Investigate Leukocyte Recruitment on Cultured Vascular Cells and Adherent Platelets

Tanja Vajen¹, Alexandra C.A. Heinzmann¹, Annemiek Dickhout¹, Zhen Zhao², Magdolna Nagy¹, Johan W.M. Heemskerk¹, Rory R. Koenen¹

¹Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, ²Department of Vascular Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine

Leukocytes avidly interact with vascular cells and platelets after vessel wall injury or during inflammation. Here, we describe a straightforward laminar flow-based assay to characterize the molecular mechanisms that underlie the interactions between leukocytes and their cellular partners.

Behavior

Investigating Pain-Related Avoidance Behavior using a Robotic Arm-Reaching Paradigm

Eveliina Glogan^1,2, Rena Gatzounis¹, Kristof Vandael^1,3, Mathijs Franssen², Johan W. S. Vlaeyen^1,2, Ann Meulders^1,2

¹Experimental Health Psychology, Maastricht University, ²Research Group Health Psychology, KU Leuven, ³Laboratory of Biological Psychology, KU Leuven

Avoidance is central to chronic pain disability, yet adequate paradigms for examining pain-related avoidance are lacking. Therefore, we developed a paradigm that allows investigating how pain-related avoidance behavior is learned (acquisition), spreads to other stimuli (generalization), can be mitigated (extinction), and how it may subsequently re-emerge (spontaneous recovery).

Biochemistry

A Semi-Automated and Reproducible Biological-Based Method to Quantify Calcium Deposition In Vitro

Armand M. G. Jaminon^*1, Nikolas Rapp^*1, Asim C. Akbulut¹, Robert Dzhanaev², Chris P. Reutelingsperger¹, Willi Jahnen-Dechent², Leon J. Schurgers^1,3

¹Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, ²Helmholtz Institute for Biomedical Engineering, Biointerface Group, RWTH Aachen University, ³Institute of Experimental Medicine and Systems Biology, RWTH Aachen University

Cardiovascular disease is the leading cause of death worldwide. Vascular calcification contributes substantially to the burden of cardiovascular morbidity and mortality. This protocol describes a simple method to quantify vascular smooth muscle cell-mediated calcium precipitation in vitro by fluorescent imaging.

Medicine

A Minimally Invasive Model of Aortic Stenosis in Swine

Rui Cerqueira^*1,2, Liliana Moreira-Costa^*1, Evangelia Beslika^1,3, André Leite-Moreira¹, Joana Silva^3,4, Paula A. da Costa Martins³, Adelino Leite-Moreira^1,2, André Lourenço¹, Pedro Mendes-Ferreira¹

¹UnIC@RISE, Department of Surgery and Physiology, Faculty of Medicine, University of Porto, ²Department of Cardiothoracic Surgery, Hospital Universitário de São João, ³CARIM School for Cardiovascular Diseases, Maastricht University, ⁴Mirabilis Therapeutics

This protocol describes a minimally invasive surgical procedure for ascending aortic banding in swine.