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University of Miami Leonard M. Miller School of Medicine

5 ARTICLES PUBLISHED IN JoVE

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JoVE Journal

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET
Alexander Grabner *1, Dominik Kentrup *1, Uta Schnöckel 2, Gert Gabriëls 1, Rita Schröter 1, Hermann Pavenstädt 1, Otmar Schober 2, Eberhard Schlatter 1, Michael Schäfers *3, Stefan Reuter *1
1Department of Internal Medicine D, Experimental Nephrology, University of Münster, 2Department of Nuclear Medicine, University of Münster, 3European Institute for Molecular Imaging, University of Münster

We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.

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Immunology and Infection

Induction of an Inflammatory Response in Primary Hepatocyte Cultures from Mice
Brian Czaya *1,2, Saurav Singh *1,2, Christopher Yanucil 1,2, Karla Schramm 1,2, Christian Faul 1,2, Alexander Grabner 1
1Katz Family Drug Discovery Center and Division of Nephrology and Hypertension, Department of Medicine, University of Miami Leonard M. Miller School of Medicine, 2Department of Cell Biology and Anatomy, University of Miami Leonard M. Miller School of Medicine

Here, we show an enzymatic approach to isolate primary hepatocytes from adult mice, and we describe the quantification of an inflammatory response using ELISA and real-time PCR.

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Biology

Isolation, Characterization, And High Throughput Extracellular Flux Analysis of Mouse Primary Renal Tubular Epithelial Cells
Wen Ding 1,2, Keyvan Yousefi 1,2, Lina A. Shehadeh 2,3,4,5
1Department of Molecular and Cellular Pharmacology, University of Miami Leonard M. Miller School of Medicine, 2Interdisciplinary Stem Cell Institute, University of Miami Leonard M. Miller School of Medicine, 3Department of Medicine, Division of Cardiology, University of Miami Leonard M. Miller School of Medicine, 4Vascular Biology Institute, University of Miami Leonard M. Miller School of Medicine, 5Peggy and Harold Katz Family Drug Discovery Center, University of Miami Leonard M. Miller School of Medicine

This protocol provides a cost-effective approach to isolate and characterize mouse primary renal tubular cells that can subsequently be sub-cultured to assess renal biological functions ex vivo, including mitochondrial bioenergetics.

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Biology

LDL Cholesterol Uptake Assay Using Live Cell Imaging Analysis with Cell Health Monitoring
Portia Ritter *1,2, Keyvan Yousefi *1,3, Juliana Ramirez 4, Derek M. Dykxhoorn 4,5, Armando J. Mendez 6, Lina A. Shehadeh 1,2,7,8
1Interdisciplinary Stem Cell Institute, University of Miami Leonard M. Miller School of Medicine, 2Department of Medicine, Division of Cardiology, University of Miami Leonard M. Miller School of Medicine, 3Department of Molecular and Cellular Pharmacology, University of Miami Leonard M. Miller School of Medicine, 4Dr. John T. Macdonald Foundation Department of Human Genetics, University of Miami Leonard M. Miller School of Medicine, 5John P. Hussman Institute for Human Genomics, University of Miami Leonard M. Miller School of Medicine, 6Department of Medicine, Division of Endocrinology, Metabolism and Endocrinology and the Diabetes Research Institute, University of Miami Leonard M. Miller School of Medicine, 7Vascular Biology Institute, University of Miami Leonard M. Miller School of Medicine, 8Peggy and Harold Katz Family Drug Discovery Center, University of Miami Leonard M. Miller School of Medicine

This protocol provides an efficient approach to measuring LDL cholesterol uptake with real time influx rates using a live cell imaging system in various cell types. This technique provides a platform to screen the pharmacological activity of compounds affecting LDL influx while monitoring for cell morphology and hence potential cytotoxicity.

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Medicine

A Murine Model of Hyperlipidemia-Induced Heart Failure with Preserved Ejection Fraction
Monique Williams 1,2, Ali Kamiar 2,3, Jose Manuel Condor Capcha 1,2, Monica Anne Rasmussen 4, Qusai Alitter 2, Rosemeire Kanashiro Takeuchi 2,3, Lauro Mitsuru Takeuchi 2, Joshua M. Hare 1,2, Lina A. Shehadeh 1,2
1Department of Medicine, Division of Cardiology, University of Miami Leonard M. Miller School of Medicine, 2Interdisciplinary Stem Cell Institute, University of Miami Leonard M. Miller School of Medicine, 3Department of Molecular and Cellular Pharmacology, University of Miami Leonard M. Miller School of Medicine, 4Department of Medical Education, University of Miami Leonard M. Miller School of Medicine

This protocol presents a detailed approach to replicating a murine model of hyperlipidemia-induced heart failure with preserved ejection fraction (HFpEF). The design combines the administration of adeno-associated virus 9-cardiac troponin T-low-density lipoprotein receptor (AAV9-cTnT-LDLR) and poloxamer-407 (P-407).

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