Wake Forest Baptist Medical Center

3 ARTICLES PUBLISHED IN JoVE

Biology

A Faster, High Resolution, mtPA-GFP-based Mitochondrial Fusion Assay Acquiring Kinetic Data of Multiple Cells in Parallel Using Confocal Microscopy

Alenka Lovy¹, Anthony J.A. Molina², Fernanda M. Cerqueira³, Kyle Trudeau³, Orian S. Shirihai³

¹Department of Neuroscience, Center for Neuroscience Research, Tufts School of Medicine, ²Department of Internal Medicine, Geriatrics & Gerontology, Wake Forest Baptist Medical Center, ³Department of Medicine, Boston University Medical Center

Mitochondrial fusion was measured by tracking the equilibration of photoconverted matrix-targeted GFP across the mitochondrial network over time. Thus far, only one cell could be subjected to an hour long kinetic analysis at a time. We present a method that simultaneously measures multiple cells, thereby speeding up the data collection process.

Bioengineering

Detergent-Free Decellularization of the Human Pancreas for Soluble Extracellular Matrix (ECM) Production

Riccardo Tamburrini^1,2,3, Deborah Chaimov^1,3, Amish Asthana^1,3, Carlo Gazia^3,4, Kevin Enck³, Sean M. Muir⁵, Justine Mariam Aziz⁶, Sandrine Lablanche⁷, Emily Tubbs⁷, Alice A. Tomei^8,9, Mark Van Dyke¹⁰, Shay Soker³, Emmanuel C. Opara³, Giuseppe Orlando^1,3

¹Department of Surgery, Wake Forest Baptist Medical Center, ²Department of General Surgery, PhD Program in Experimental Medicine, University of Pavia, ³Wake Forest Institute for Regenerative Medicine, Wake Forest School of Medicine, ⁴Department of Surgery, Tor Vergata University of Rome, ⁵Wake Forest University College of Arts and Science, ⁶Wake Forest University School of Medicine, ⁷Laboratory of Fundamental and Applied Bioenergetics (LBFA), and Environmental and System Biology (BEeSy), Grenoble Alps University, ⁸Department of Biomedical Engineering, University of Miami, ⁹Diabetes Research Institute, University of Miami Miller School of Medicine, ¹⁰Department of Biomedical Engineering and Mechanics, School of Biomedical Engineering and Sciences, Virginia Polytechnic Institute and State University

The protocol described in this manuscript explains the steps for the fabrication of a soluble extracellular matrix (ECM) from the human pancreas. The solubilized ECM powder obtained through this protocol may be used for the recapitulation of pancreatic islets’ microenvironment in vitro and, potentially, in vivo settings.

Immunology and Infection

Retrograde Parotid Gland Infusion through Stensen's Duct in a Non-Human Primate for Vectored Gene Delivery

Guy El Helou¹, Joseph F. Goodman², Maria Blevins³, David L. Caudell⁴, Todd A. Ponzio³, John W. Sanders^3,5

¹Department of Medicine, Division of Infectious Diseases and Global Medicine, University of Florida, ²Department of Otolaryngology, George Washington University School of Medicine, ³Department of Medicine, Section on Infectious Diseases, Wake Forest Baptist Medical Center, ⁴Department of Pathology, Wake Forest School of Medicine, ⁵Department of Medicine, Section of Infectious Diseases, Hefner Veterans Affairs Medical Center

Salivary glands have been proposed as a tissue target site for gene therapy, especially in the area of vaccination by gene transfer. We demonstrate gene delivery in a non-human primate model utilizing retrograde parotid infusion.