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Tulane University

17 ARTICLES PUBLISHED IN JoVE

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Medicine

An Orthotopic Model of Murine Bladder Cancer
Georgina L. Dobek 1, W. T. Godbey 2
1Department of Comparative Medicine, Tulane University, 2Department of Chemical and Biomolecular Engineering, Tulane University

Bladder tumors are established in female mice in a minimally invasive fashion through catheterization, local cauterization, and subsequent adhesion of carcinoma cells to the burn sites.

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Bioengineering

Fabrication of Micropatterned Hydrogels for Neural Culture Systems using Dynamic Mask Projection Photolithography
J. Lowry Curley 1, Scott R. Jennings 1, Michael J. Moore 1
1Biomedical Engineering, Tulane University

Simple techniques are described for the rapid production of microfabricated neural culture systems using a digital micromirror device for dynamic mask projection lithography on regular cell culture substrates. These culture systems may be more representative of natural biological architecture, and the techniques described could be adapted for numerous applications.

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Biology

Rat Mesentery Exteriorization: A Model for Investigating the Cellular Dynamics Involved in Angiogenesis
Ming Yang 1, Peter C. Stapor 1, Shayn M. Peirce 2, Aline M. Betancourt 3, Walter L. Murfee 1
1Department of Biomedical Engineering, Tulane University, 2Department of Biomedical Engineering, University of Virginia , 3Center for Stem Cell Research and Regenerative Medicine, Tulane University

This article describes a simple model for stimulating angiogenesis in the rat mesentery. The model produces dramatic increases in capillary sprouting, vascular area and vascular density over a relatively short time course in a tissue that allows en face visualization of entire microvascular networks down to the single cell level.

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Bioengineering

Nonhuman Primate Lung Decellularization and Recellularization Using a Specialized Large-organ Bioreactor
Ryan W. Bonvillain 1,2, Michelle E. Scarritt 1, Nicholas C. Pashos 1, Jacques P. Mayeux 1, Christopher L. Meshberger 1, Aline M. Betancourt 1,3, Deborah E. Sullivan 1,3, Bruce A. Bunnell 1,2,4
1Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, 2Division of Regenerative Medicine, Tulane National Primate Research Center, 3Department of Microbiology and Immunology, Tulane University School of Medicine, 4Department of Pharmacology, Tulane University School of Medicine

Whole-organ decellularization produces natural biological scaffolds that may be used for regenerative medicine. The description of a nonhuman primate model of lung regeneration in which whole lungs are decellularized and then seeded with adult stem cells and endothelial cells in a bioreactor that facilitates vascular circulation and liquid media ventilation is presented.

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Medicine

An Alkali-burn Injury Model of Corneal Neovascularization in the Mouse
Chastain Anderson *1, Qinbo Zhou *1, Shusheng Wang 1,2
1Department of Cell and Molecular Biology, Tulane University, 2Department of Ophthalmology, Tulane University

Neovascularization (NV) of the cornea can complicate multiple visual pathologies. Utilizing a controlled, alkali-burn injury model, a quantifiable level of corneal NV can be produced for mechanistic study of corneal NV and evaluation of potential therapies for neovascular disorders.

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Developmental Biology

An Ex Vivo Method for Time-Lapse Imaging of Cultured Rat Mesenteric Microvascular Networks
Mohammad S. Azimi 1, Jessica M. Motherwell 1, Walter L. Murfee 1
1Biomedical Engineering, Tulane University

Angiogenesis involves multi-cell, multi-system interactions that need to be investigated in a physiologically relevant environment. The objective of this study is to demonstrate the ability of the rat mesentery culture model to make time-lapse comparisons of intact microvascular networks during angiogenesis.

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Immunology and Infection

Phenotypic Analysis of Rodent Malaria Parasite Asexual and Sexual Blood Stages and Mosquito Stages
Ahmed S.I. Aly 1,2, Gozde Deveci 1, Ilknur Yilmaz 1, Amanah Abraham 2, Aneesa Golshan 2, Robert J. Hart 2
1Beykoz Institute of Life Sciences and Biotechnology, Bezmialem Vakif University, Istanbul, Turkey, 2School of Public Health and Tropical Medicine, Department of Tropical Medicine, Tulane University

Due to the striking similarities of the life cycle and biology of rodent malaria parasites to human malaria parasites, rodent malaria models have become indispensable for malaria research. Herein, we standardized some of the most important techniques used in the phenotypic analysis of wild-type and transgenic rodent malaria species.

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Bioengineering

A Microcontroller Operated Device for the Generation of Liquid Extracts from Conventional Cigarette Smoke and Electronic Cigarette Aerosol
Chastain A. Anderson 1, Rachael E. Bokota 1, Andrew E. Majeste 1, Walter L. Murfee 2, Shusheng Wang 3
1Cell & Molecular Biology, Tulane University, 2Biomedical Engineering, Tulane University, 3Cell & Molecular Biology & Ophthalmology, Tulane University

Here, we describe a programmable laboratory device that can be used to create extracts of conventional cigarette smoke and electronic cigarette aerosol. This method provides a useful tool for making direct comparisons between conventional cigarettes and electronic cigarettes, and is an accessible entry point into electronic cigarette research.

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JoVE Core

Characterization of Synthetic Polymers via Matrix Assisted Laser Desorption Ionization Time of Flight (MALDI-TOF) Mass Spectrometry
Molly E. Payne 1, Scott M. Grayson 1
1Department of Chemistry, Tulane University

A protocol for the matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) characterization of synthetic polymers is described including the optimization of sample preparation, spectral acquisition, and data analysis.

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JoVE Journal

RNA Next-Generation Sequencing and a Bioinformatics Pipeline to Identify Expressed LINE-1s at the Locus-Specific Level
Tiffany Kaul 1, Maria E. Morales 1, Emily Smither 1, Melody Baddoo 1,2, Victoria P. Belancio 1,3, Prescott Deininger 1,4
1Tulane Cancer Center, Tulane University, 2Department of Pathology, Tulane University, 3Department of Structural and Cellular Biology, Tulane University, 4Department of Epidemiology, Tulane University

Here, we present a bioinformatic approach and analyses to identify LINE-1 expression at the locus specific level.

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Bioengineering

Biaxial Basal Tone and Passive Testing of the Murine Reproductive System Using a Pressure Myograph
Shelby E. White 1, Cassandra K. Conway 1, Gabrielle L. Clark 1, Dylan J. Lawrence 1, Carolyn L. Bayer 1, Kristin S. Miller 1
1Department of Biomedical Engineering, Tulane University

This protocol utilized a commercially available pressure myograph system to perform pressure myograph testing on the murine vagina and cervix. Utilizing media with and without calcium, the contributions of the smooth muscle cells (SMC) basal tone and passive extracellular matrix (ECM) were isolated for the organs under estimated physiological conditions.

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Medicine

A Rat Model of Pressure Overload Induced Moderate Remodeling and Systolic Dysfunction as Opposed to Overt Systolic Heart Failure
Antoine H. Chaanine 1, L. Gabriel Navar 2, Patrice Delafontaine 1,2,3
1Department of Medicine/Heart and Vascular Institute, Tulane University, 2Department of Physiology, Tulane University, 3Department of Pharmacology, Tulane University

We describe the creation of a rat model of pressure overload induced moderate remodeling and early systolic dysfunction where signal transduction pathways involved in the initiation of the remodeling process are activated. This animal model will aid in identifying molecular targets for applying early therapeutic anti-remodeling strategies for heart failure.

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Behavior

A Novel Pavlovian Fear Conditioning Paradigm to Study Freezing and Flight Behavior
Chandrashekhar D. Borkar 1,2, Jonathan P. Fadok 1,2
1Department of Psychology, Tulane University, 2Tulane Brain Institute, Tulane University

Defensive behavioral responses are contingent upon threat intensity, proximity, and context of exposure. Based on these factors, we developed a classical conditioning paradigm that elicits clear transitions between conditioned freezing and flight behavior within individual subjects. This model is crucial for the understanding the pathologies involved in anxiety, panic, and post-traumatic stress disorders.

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Cancer Research

Modeling Breast Cancer in Human Breast Tissue using a Microphysiological System
Loren M. Brown 1, Katherine L. Hebert 2, Rakesh R. Gurrala 3, C. Ethan Byrne 4, Matthew Burow 5, Elizabeth C. Martin 4, Frank H. Lau 1
1Department of Surgery, Louisiana State University Health Sciences Center, 2Department of Bioinnovation, Tulane University, 3Tulane University School of Medicine, 4Department of Biological and Agricultural Engineering, Louisiana State University, 5Department of Medicine, Tulane University School of Medicine

This protocol describes the construction of an in vitro microphysiological system for studying breast cancer using primary human breast tissue with off the shelf materials.

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Bioengineering

Earthworm, Lumbricus Terrestris: A Novel Microinjection Vasculature In vivo Invertebrate Model
Asis Lopez 1,5, Yaswitha Mikkilineni 2, Shayna Berman 3, Damir Khismatullin 1,4, Gregory T. Clement 5, Matthew R. Myers 5
1Bioinnovation PhD Program, Tulane University, 2Biology Department, Virginia Commonwealth University, 3Mechanical Engineering Department, Montgomery College, 4Biomedical Engineering Department, Tulane University, 5US Food and Drug Administration

Earthworms are a novel invertebrate in vivo bench-top model for vasculature studies. We present techniques and equipment that allow efficient surgery and microinjection into the earthworm vasculature. Surgical protocols, microinjection techniques and the procedure for producing custom-made micropipettes are described.

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Bioengineering

Preparation and Structural Evaluation of Epithelial Cell Monolayers in a Physiologically Sized Microfluidic Culture Device
Eshan B. Damle 1, Eiichiro Yamaguchi 1, Joshua E. Yao 1, Donald P. Gaver III 1
1Department of Biomedical Engineering, School of Science and Engineering, Tulane University

The presented protocol describes the development and use of a phalloidin-based filamentous-actin staining technique with confocal laser scanning microscopy (CLSM) to visualize adherent cell layer structure in microfluidic dynamic-culture channels and traditional fixed-well static-culture chambers. This approach aids in evaluating cell layer confluency, monolayer formation, and layer-thickness uniformity.

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Genetics

Monochrome Multiplex Quantitative PCR Telomere Length Measurement
Noelle A. Martin *1, Lauren W. Y. McLester-Davis *2,3,4, Trevor R. Roy 1, Madeline G. Magruder 5, Waylon J. Hastings 1, Stacy S. Drury 6
1School of Medicine, Tulane University, 2Native American Center for Health Professions, University of Wisconsin-Madison, 3Department of Medicine, University of Wisconsin-Madison, 4Department of Biochemistry, University of Wisconsin-Madison, 5School of Science and Engineering, Tulane University, 6Department of Psychiatry and Behavioral Sciences, Boston Children’s Hospital

Here we present a protocol for the measurement of relative telomere length (TL) using the monochrome multiplex quantitative polymerase chain reaction (MMqPCR) assay. The MMqPCR assay is a repeatable, efficient, and cost-effective method for measuring TL from human DNA in population-based studies.

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