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King’s College London

6 ARTICLES PUBLISHED IN JoVE

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Biology

Isolation and Culture of Neonatal Mouse Cardiomyocytes
Elisabeth Ehler 1, Thomas Moore-Morris 2, Stephan Lange 2
1Randall and Cardiovascular Divisions, King’s College London, 2Division of Cardiology, School of Medicine, University of California San Diego

Primary mouse cardiomyocyte cultures are one of the pivotal tools for the investigation of myofibrillar organization and function. The following protocol describes the isolation and culture of primary cardiomyocytes from neonatal mouse hearts. The resulting cardiomyocyte cultures may be subsequently used for a variety of biomechanical, biochemical and cell-biological assays.

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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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Immunology and Infection

Co-Culture of Murine Small Intestine Epithelial Organoids with Innate Lymphoid Cells
Emily Read *1,2, Geraldine M. Jowett *1,2,3, Diana Coman 1, Joana F. Neves 1
1Centre for Host-Microbiome Interactions, King’s College London, 2Wellcome Trust Cell Therapies and Regenerative Medicine Ph.D. Programme, King’s College London, 3Present address: Wellcome Trust/Cancer Research UK Gurdon Institute, Cambridge University

This protocol offers detailed instructions for establishing murine small intestine organoids, isolating type-1 innate lymphoid cells from the murine small intestine lamina propria, and establishing 3-dimensional (3D) co-cultures between both cell types to study bi-directional interactions between intestinal epithelial cells and type-1 innate lymphoid cells.

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Developmental Biology

Real Time and Repeated Measurement of Skeletal Muscle Growth in Individual Live Zebrafish Subjected to Altered Electrical Activity
Michael Attwaters *1, Jeffrey J. Kelu *1, Tapan G. Pipalia 1, Simon M. Hughes 1
1Randall Centre for Cell and Molecular Biophysics, School of Basic and Medical Biosciences, King’s College London

Optical clarity is a major advantage for cell biological and physiological work in zebrafish. Robust methods for measurement of cell growth in individual animals are described that permit novel insights into how growth of skeletal muscle and neighboring tissues are integrated with whole body growth.

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Biochemistry

Optimized Methods for the Surface Immobilization of Collagens and Collagen Binding Assays
Nadia Chaher 1, Giuseppe Digilio *2, Sara Lacerda *3, René M. Botnar 1,4,5,6, Alkystis Phinikaridou 1,4
1School of Biomedical Engineering and Imaging Sciences, King’s College London, 2Dipartimento di Scienze e Innovazione Tecnologica, Università del Piemonte Orientale, 3Centre de Biophysique Moléculaire, CNRS UPR 4301, Université d'Orléans, 4BHF Centre of Research Excellence, Cardiovascular Division, King’s College London, 5Escuela de Ingeniería, Pontificia Universidad Católica de Chile, 6Instituto de Ingeniería Biológica y Médica, Pontificia Universidad Católica de Chile

This work presents an optimized protocol to reproducibly immobilize and quantify type I and III collagen onto microplates, followed by an improved in vitro binding assay protocol to study collagen-compound interactions using a time-resolved fluorescence method. The subsequent step-by-step data analysis and data interpretation are provided.

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Recent Advances and Best Practices for Cryo-EM Analysis for Protein Structure Determination
Julien Bergeron 1, Chiara Rapisarda 2
1Randall Centre for Cell and Molecular Biophysics, King’s College London, 2Bio Structure and Biophysics (BSB), Sanofi

Recent Advances and Best Practices for Cryo-EM Analysis for Protein Structure Determination

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