City of Hope View Institution's Website 5 articles published in JoVE Medicine Isolation and Profiling of Human Primary Mesenteric Arterial Endothelial Cells at the Transcriptome Level Naseeb Kaur Malhi*1, Yingjun Luo*1, Xiaofang Tang*1, Kiran Sriram1,3, Riccardo Calandrelli2, Sheng Zhong2, Zhen Bouman Chen1,3 1Department of Diabetes Complications and Metabolism, City of Hope, 2Department of Bioengineering, University of California San Diego, 3Irell and Manella Graduate School of Biological Sciences, City of Hope The protocol describes the isolation, culture, and profiling of endothelial cells from human mesenteric artery. Additionally, a method is provided to prepare human artery for spatial transcriptomics. Proteomics, transcriptomics, and functional assays can be performed on isolated cells. This protocol can be repurposed for any medium- or large-size artery. Biology A Quantitative Assay for Insulin-expressing Colony-forming Progenitors Michael Winkler1, Nancy Trieu2, Tao Feng2, Liang Jin2, Stephanie Walker2, Lipi Singh2, Hsun Teresa Ku2,3 1Department of Biotechnology & Bioinformatics, California State University Channel Islands, 2Department of Diabetes, Endocrinology and Metabolism, Beckman Research Institute of City of Hope, 3The Irell & Manella Graduate School of Biological Sciences, Beckman Research Institute of City of Hope A three-dimensional clonogenic assay that allows pancreatic-like progenitors to differentiate into insulin-expressing colonies is described. This method takes advantage of semi-solid media containing methylcellulose, Matrigel and growth factors, in which single progenitors proliferate and differentiate in vitro, permitting quantification of the number of functional progenitors in a population. Biology Quantitation and Analysis of the Formation of HO-Endonuclease Stimulated Chromosomal Translocations by Single-Strand Annealing in Saccharomyces cerevisiae Lauren Liddell1,2, Glenn Manthey2, Nicholas Pannunzio3, Adam Bailis2 1Irell & Manella Graduate School of Biological Sciences, 2Department of Molecular and Cellular Biology, City of Hope Comprehensive Cancer Center and Beckman Research Institute, 3Department of Biochemistry and Molecular Biology, University of Southern California, Norris Comprehensive Cancer Center The HO-stimulated translocation assay monitors single-strand annealing following the creation of DNA double-strand breaks at multiple loci in diploid Saccharomyces cerevisiae. This mechanism may model genome rearrangements in somatic cells of higher eukaryotes following exposure to high doses of ionizing radiation. Immunology and Infection Development of Cell-type specific anti-HIV gp120 aptamers for siRNA delivery Jiehua Zhou1, Haitang Li1, Jane Zhang2, Swiderski Piotr3, John Rossi1 1Department of Molecular and Cellular Biology, Beckman Research Institute of City of Hope, 2Graduate School of Biological Sciences, Beckman Research Institute of City of Hope, 3Shared Resource-DNA/RNA Peptide, Beckman Research Institute of City of Hope Several 2’-Fluoro RNA aptamers against HIV-1Ba-L gp120 with nanomole affinity are isolated from a RNA library by in vitro SELEX procedure. A new dual inhibitory function anti-gp120 aptamer-siRNA chimera is created and shows considerable promise for systemic anti-HIV therapy. Biology Isolation and Enrichment of Rat Mesenchymal Stem Cells (MSCs) and Separation of Single-colony Derived MSCs Linxia Zhang1, Christina Chan1 1Department of Chemical Engineering and Materials Science, City of Hope Cancer Center Rat MSCs were isolated from femurs and tibias and then enriched by magnetic cell sorting. Sorted cells were confirmed for the expression of surface markers by flow cytometry. These cells were also cultured at clonal density to form single colonies and then these colonies were separated by cloning cylinders.