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Karolinska University Hospital

10 ARTICLES PUBLISHED IN JoVE

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Medicine

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time
Midhat H. Abdulreda 1,2, Alejandro Caicedo 1,3,4, Per-Olof Berggren 1,5
1Diabetes Research Institute, University of Miami Miller School of Medicine, 2Department of Surgery, University of Miami Miller School of Medicine, 3Department of Medicine, University of Miami Miller School of Medicine, 4Department of Physiology & Biophysics, University of Miami Miller School of Medicine, 5The Rolf Luft Research Center for Diabetes and Endocrinology, Karolinska Institutet

A new approach combining intraocular transplantation and confocal microscopy enables longitudinal, non-invasive real-time imaging with single-cell resolution within grafted tissues in vivo. We demonstrate how to transplant pancreatic islets into the anterior chamber of the mouse eye.

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Bioengineering

Minced Tissue in Compressed Collagen: A Cell-containing Biotransplant for Single-staged Reconstructive Repair
Clara I. Chamorro 1, Said Zeiai 1,2, Gisela Reinfeldt Engberg 1,2, Magdalena Fossum 1,2
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.

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Immunology and Infection

Increased Recovery Time and Decreased LPS Administration to Study the Vagus Nerve Stimulation Mechanisms in Limited Inflammatory Responses
Erwan Le Maître 1, Priya Revathikumar 1, Johanna Estelius 1, Jon Lampa 1
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.

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Biology

Co-immunoprecipitation Assay Using Endogenous Nuclear Proteins from Cells Cultured Under Hypoxic Conditions
Xiaofeng Zheng 1,2, Calvin Qing Wei Ho 1, Xiaowei Zheng 3, Kian Leong Lee 4, Katarina Gradin 5, Teresa S. Pereira 3, Per-Olof Berggren 1,2,3, Yusuf Ali 1,2
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.

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Developmental Biology

Clonal Genetic Tracing using the Confetti Mouse to Study Mineralized Tissues
Baoyi Zhou 1, Marketa Kaucka 1, Andrei S. Chagin 1,2, Phillip T. Newton 1,3
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.

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Biochemistry

Studies of Chaperone-Cochaperone Interactions using Homogenous Bead-Based Assay
Lisha Wang 1, Liza Bergkvist 1, Rajnish Kumar 1,2, Bengt Winblad 1,3, Pavel F. Pavlov 1
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.

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Pedicle Screw Placement Using an Augmented Reality Head-Mounted Display in a Porcine Model

Pedicle Screw Placement Using an Augmented Reality Head-Mounted Display in a Porcine Model
Henrik Frisk 1, Gustav Burström 1, Juliane Weinzierl 2, Linda Westernhagen 2, Florentin Tranchant 2, Erik Edström 1,3, Adrian Elmi-Terander 1,3
1Department of Clinical Neuroscience, Karolinska Institutet, 2Brainlab AG, 3Capio Spine Center Stockholm, Löwenströmska Hospital

The augmented reality head-mounted display, Magic Leap, was used in combination with a conventional navigation system to place pedicle screws in a porcine model by adhering to a novel workflow. With a median insertion time of <2.5 min, submillimeter technical accuracy and 100% clinical accuracy were achieved according to Gertzbein.

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Medicine

Measurement of Healthy and Injured Triceps Surae Morphology
Hayley Powell Smitheman 1, Kayla D. Seymore 1, Morgan N. Potter 1, Andy K. Smith 1, Susanna Aufwerber 2,3, Karin Grävare Silbernagel 1
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.

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Biology

In Vivo Imaging of Liver Spheroids Engrafted in the Anterior Chamber of the Mouse Eye
Francesca Lazzeri-Barcelo 1, Pierpaolo Ciardo 1, Barbara Leibiger 1, Ingo B. Leibiger 1, 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.

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Developmental Biology

Z-Scores for Assessing Ovarian Reserve in Young Patients Undergoing Fertility Preservation
Jasmin Hassan 1,2, Pauliina Damdimopoulou 1,2, Atte Lahtinen 3,4, Kirsi Jahnukainen 5,6, Katri Knuus 7, Ganesh Acharya 1, Richelle D. Björvang 1,8,9
1Division of Obstetrics and Gynecology, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, 2Department of Gynecology and Reproductive Medicine, Karolinska University Hospital, 3Applied Tumor Genomics Research Program, Faculty of Medicine, University of Helsinki, 4Department of Medical and Clinical Genetics, Medicum, Faculty of Medicine, University of Helsinki, 5Children's Hospital, Pediatric Research Centre, University of Helsinki and Helsinki University Hospital, 6NORDFERTIL Research Lab Stockholm, Department of Women's, and Children's Health, Karolinska Institutet, and Karolinska University Hospital, 7Department of Obstetrics and Gynecology, Helsinki University Hospital, University of Helsinki, 8Department of Cell and Molecular Biology, Uppsala University, 9Department of Women's and Children's Health, Uppsala University

The present protocol describes a method for assessing ovarian reserve in patients under 25 years old who require fertility preservation through ovarian tissue cryopreservation. This method involves: (1) histological assessment of ovarian reserve in cortical samples, (2) comparison to a reference dataset, and (3) calculation of Z-scores.

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