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University of Miami Leonard M. Miller School of Medicine

5 ARTICLES PUBLISHED IN JoVE

JoVE Journal

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using ¹⁸F-FDG PET

Alexander Grabner^*1, Dominik Kentrup^*1, Uta Schnöckel², Gert Gabriëls¹, Rita Schröter¹, Hermann Pavenstädt¹, Otmar Schober², Eberhard Schlatter¹, 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.

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 Grabner¹

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.

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.

Biology

LDL Cholesterol Uptake Assay Using Live Cell Imaging Analysis with Cell Health Monitoring

Portia Ritter^*1,2, Keyvan Yousefi^*1,3, Juliana Ramirez⁴, Derek M. Dykxhoorn^4,5, Armando J. Mendez⁶, Lina A. Shehadeh^1,2,7,8

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.

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 Rasmussen⁴, Qusai Alitter², Rosemeire Kanashiro Takeuchi^2,3, Lauro Mitsuru Takeuchi², 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).