Karolinska University Hospital 9 articles published in JoVE Biology In Vivo Imaging of Liver Spheroids Engrafted in the Anterior Chamber of the Mouse Eye Francesca Lazzeri-Barcelo1, Pierpaolo Ciardo1, Barbara Leibiger1, Ingo B. Leibiger1, Per-Olof Berggren*1,2,3, Noah Moruzzi*1 1The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Karolinska University Hospital, 2Tecnológico de Monterrey, 3Diabetes Research Institute, Miller School of Medicine, University of Miami Here, we describe a platform that allows noninvasive in vivo imaging of liver spheroids engrafted in the anterior chamber of the mouse eye. The workflow spans from generating spheroids from primary liver cells to transplantation into the mouse eye and in vivo imaging at cellular resolution by confocal microscopy. Medicine Measurement of Healthy and Injured Triceps Surae Morphology Hayley Powell Smitheman1, Kayla D. Seymore1, Morgan N. Potter1, Andy K. Smith1, Susanna Aufwerber2,3, Karin Grävare Silbernagel1 1Department of Physical Therapy, University of Delaware, 2Department of Molecular Medicine and Surgery, Karolinska Institute, 3Women’s Health and Allied Health Professionals Theme, Medical Unit of Occupational Therapy and Physiotherapy, Karolinska University Hospital Ultrasound imaging is becoming more accessible in clinical and research settings, and a consistent protocol will be beneficial for comparison between studies and for clinical interpretations. This protocol for ultrasound evaluation is a valid and reliable method to evaluate Achilles tendon morphology in healthy, tendinopathic, and ruptured tendons. Biochemistry Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay Lisha Wang1, Liza Bergkvist1, Rajnish Kumar1,2, Bengt Winblad1,3, Pavel F. Pavlov1 1Department of Neuroscience Care and Society, Division of Neurogeriatrics, Karolinska Institutet, 2Department of Pharmaceutical Engineering & Technology, Indian Institute of Technology (BHU), 3Theme Inflammation and Aging, Karolinska University Hospital This protocol presents a technique for probing protein-protein interactions using glutathione-linked donor beads with GST-fused TPR-motif co-chaperones and acceptor beads coupled with an Hsp90-derived peptide. We have used this technique to screen small molecules to disrupt Hsp90-FKBP51 or Hsp90-FKBP52 interactions and identified potent and selective Hsp90-FKBP51 interaction inhibitors. Developmental Biology Clonal Genetic Tracing using the Confetti Mouse to Study Mineralized Tissues Baoyi Zhou1, Marketa Kaucka1, Andrei S. Chagin1,2, Phillip T. Newton1,3 1Department of Physiology and Pharmacology, Karolinska Institutet, 2Institute for Regenerative Medicine, Sechenov First Moscow State Medical University (Sechenov University), 3Department of Women's and Children's Health, Karolinska Institutet and Pediatric Endocrinology Unit, Karolinska University Hospital This method describes the use of the R26R-Confetti (Confetti) mouse model to study mineralized tissues, covering all steps from the breeding strategy to the image acquirements. Included is a general protocol that can be applied to all soft tissues and a modified protocol that can be applied to mineralized tissues. Immunology and Infection Quantification of Atherosclerosis in Mice Monica Centa1, Daniel F.J. Ketelhuth1,2, Stephen Malin1, Anton Gisterå1 1Cardiovascular Medicine Unit, Center for Molecular Medicine, Department of Medicine, Karolinska Institutet, Karolinska University Hospital, 2Department of Cardiovascular and Renal Research, Institute for Molecular Medicine, University of Southern Denmark (SDU) Murine models of atherosclerosis are useful tools to investigate pathogenic pathways on a molecular level, but require standardized quantification of lesion development. This protocol describes an optimized method to determine lesion size in the major arterial vessels including the aortic root, aortic arch, and brachiocephalic artery. Biology Co-immunoprecipitation Assay Using Endogenous Nuclear Proteins from Cells Cultured Under Hypoxic Conditions Xiaofeng Zheng1,2, Calvin Qing Wei Ho1, Xiaowei Zheng3, Kian Leong Lee4, Katarina Gradin5, Teresa S. Pereira3, Per-Olof Berggren1,2,3, Yusuf Ali1,2 1Lee Kong Chian School of Medicine, Nanyang Technological University, 2Singapore Eye Research Institute (SERI), Singapore General Hospital, 3The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet, Karolinska University Hospital, 4Cancer and Stem Cell Biology Program, Duke-NUS Medical School, 5Department of Cell and Molecular Biology, Karolinska Institutet Here we describe a co-immunoprecipitation protocol to study protein-protein interactions between endogenous nuclear proteins under hypoxic conditions. This method is suitable for demonstration of the interactions between transcription factors and transcriptional co-regulators at hypoxia. Immunology and Infection Assessment of the Synaptic Interface of Primary Human T Cells from Peripheral Blood and Lymphoid Tissue Maria Steblyanko1, Nadia Anikeeva1, Marcus Buggert2,3, Michael R. Betts2, Yuri Sykulev4 1Department of Microbiology and Immunology, Thomas Jefferson University, 2Department of Microbiology and Institute for Immunology, Perelman School of Medicine, University of Pennsylvania, 3Department of Medicine Huddinge, Karolinska Institutet, Karolinska University Hospital Huddinge, 4Departments of Microbiology and Immunology and Medical Oncology, Sidney Kimmel Cancer Center, Thomas Jefferson University The protocol describes a technique to study the ability of primary polyclonal human T cells to form synaptic interfaces using planar lipid bilayers. We use this technique to show the differential synapse formation capability of human primary T cells derived from lymph nodes and peripheral blood. Immunology and Infection Increased Recovery Time and Decreased LPS Administration to Study the Vagus Nerve Stimulation Mechanisms in Limited Inflammatory Responses Erwan Le Maître1, Priya Revathikumar1, Johanna Estelius1, Jon Lampa1 1Department of Medicine, Unit of Rheumatology, Center for Molecular Medicine (CMM), Karolinska Institute, Karolinska University Hospital Vagus nerve stimulation has proven to have a strong efficacy for decreasing peripheral inflammation. Here, we present a modified vagus nerve stimulation protocol that allows for further examinations of the cholinergic anti-inflammatory mechanisms in limited inflammatory responses. Bioengineering Minced Tissue in Compressed Collagen: A Cell-containing Biotransplant for Single-staged Reconstructive Repair Clara I. Chamorro1, Said Zeiai1,2, Gisela Reinfeldt Engberg1,2, Magdalena Fossum1,2 1Department of Women's and Children's Health, Center for Molecular Medicine, Karolinska Institutet, 2Department of Pediatric Surgery, Urology Section, Astrid Lindgren Children's Hospital, Karolinska University Hospital Tissue engineering often includes in vitro expansion in order to create autografts for tissue regeneration. In this study a method for tissue expansion, regeneration, and reconstruction in vivo was developed in order to minimize the processing of cells and biological materials outside the body.
