Rutgers Robert Wood Johnson Medical School

6 ARTICLES PUBLISHED IN JoVE

Medicine

Laser-capture Microdissection of Human Prostatic Epithelium for RNA Analysis

Giovanni Lugli¹, Yachana Kataria¹, Zachary Richards¹, Peter Gann¹, Xiaofeng Zhou², Larisa Nonn¹

¹Department of Pathology, University of Illinois at Chicago, ²Department of Periodontics, University of Illinois at Chicago

The goal of this protocol is to use laser-capture micro-dissection as an effective method to isolate pure populations of cell types from heterogeneous prostate tissues for downstream RNA analysis.

Neuroscience

Quantitative Measurement of Relative Retinoic Acid Levels in E8.5 Embryos and Neurosphere Cultures Using the F9 RARE-Lacz Cell-based Reporter Assay

Myka R. Ababon^*1, Bo I. Li^*2, Paul G. Matteson¹, James H. Millonig^1,2

¹Center for Advanced Biotechnology and Medicine, Rutgers University, ²Department of Neuroscience and Cell Biology, Rutgers University

Methods to accurately measure retinoic acid (RA) levels in small amounts of tissue do not exist. This protocol describes the easy, quantitative measurement of relative RA levels in E8.5 embryos and neurospheres using an RA reporter cell line.

Developmental Biology

Rapid Detection of Neurodevelopmental Phenotypes in Human Neural Precursor Cells (NPCs)

Madeline Williams^*1, Smrithi Prem^*1, Xiaofeng Zhou¹, Paul Matteson², Percy Luk Yeung³, Chi-Wei Lu³, Zhiping Pang⁴, Linda Brzustowicz⁵, James H. Millonig², Emanuel Dicicco-Bloom¹

¹Department of Neuroscience and Cell Biology, Rutgers Robert Wood Johnson Medical School, ²Center for Advanced Biotechnology and Medicine, Department of Neuroscience and Cell Biology, Rutgers Robert Wood Johnson Medical School, ³The Child Health Institute of NJ, Department of Obstetrics, Gynecology, and Reproductive Services, Rutgers Robert Wood Johnson Medical School, ⁴The Child Health Institute of NJ, Department of Neuroscience and Cell Biology, Rutgers Robert Wood Johnson Medical School, ⁵Department of Genetics, Rutgers University

Neurodevelopmental processes such as proliferation, migration, and neurite outgrowth are often perturbed in neuropsychiatric diseases. Thus, we present protocols to rapidly and reproducibly assess these neurodevelopmental processes in human iPSC-derived NPCs. These protocols also allow the assessment of the effects of relevant growth factors and therapeutics on NPC development.

Cancer Research

Phosphopeptide Enrichment Coupled with Label-free Quantitative Mass Spectrometry to Investigate the Phosphoproteome in Prostate Cancer

Larry C. Cheng^*1,2, Zhen Li^*3, Thomas G. Graeber⁴, Nicholas A. Graham⁵, Justin M. Drake^1,2,3,6,7

¹Graduate Program in Cellular and Molecular Pharmacology, School of Graduate Studies, Rutgers University, The State University of New Jersey, ²Graduate Program in Quantitative Biomedicine, School of Graduate Studies, Rutgers University, The State University of New Jersey, ³Department of Medicine, Division of Medical Oncology, Rutgers Robert Wood Johnson Medical School, ⁴Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, Jonsson Comprehensive Cancer Center, UCLA Metabolomics Center, and California NanoSystems Institute, David Geffen School of Medicine, University of California, Los Angeles, ⁵Mork Family Department of Chemical Engineering and Materials Science, University of Southern California, ⁶Pharmacology, Rutgers Robert Wood Johnson Medical School, ⁷Cancer Metabolism and Growth Program, Rutgers Cancer Institute of New Jersey

This protocol describes a procedure to extract and enrich phosphopeptides from prostate cancer cell lines or tissues for an analysis of the phosphoproteome via mass spectrometry-based proteomics.

Engineering

Radio Frequency Magnetron Sputtering of GdBa₂Cu₃O₇₋_δ/ La_0.67Sr_0.33MnO₃ Quasi-bilayer Films on SrTiO₃ (STO) Single-crystal Substrates

Ying Wang^1,2, Zhen Li^1,2, YongSheng Liu¹, Yijie Li², Linfei Liu², Da Xu², Xiaojing Luo¹, Tian Gao^1,3, Yanyan Zhu^1,4, Luozeng Zhou³, Jianming Xu³

¹Department of Physics, Mathematics, Shanghai Key Laboratory of Materials Protection and Advanced Materials in Electric Power, Shanghai University of Electric Power, ²Key Laboratory of Artificial Structure and Quantum Control, Ministry of Education, Department of Physics, Shanghai Jiao Tong University, ³Shanghai Institute of Space Power-sources, ⁴Shanghai Key Laboratory of High Temperature Superconductors, Shanghai University

Here, we present a protocol to grow LSMO nanoparticles and (Gd) BCO films on (001) SrTiO₃ (STO) single-crystal substrates by radio frequency (RF)-sputtering.

Medicine

A Proinflammatory, Degenerative Organ Culture Model to Simulate Early-Stage Intervertebral Disc Disease.

Babak Saravi^*1,2, Gernot Lang^*2, Sibylle Grad¹, Mauro Alini¹, R. Geoff Richards^1,2, Hagen Schmal², Norbert Südkamp², Zhen Li¹

¹AO Research Institute Davos, Davos, Switzerland, ²Department of Orthopedics and Trauma Surgery, Medical Centre-Albert-Ludwigs-University of Freiburg, Faculty of Medicine, Albert-Ludwigs-University of Freiburg, Freiburg, Germany

This protocol presents a novel experimental model of proinflammatory, degenerative bovine organ culture to simulate early-stage intervertebral disc degeneration.