Peking Union Medical College 6 articles published in JoVE Immunology and Infection Establishment of Deep Hypothermic Circulatory Arrest in Rats Weidong Yan1, Bingyang Ji1 1Department of Cardiopulmonary Bypass, National Center for Cardiovascular Disease and Fuwai Hospital, State Key Laboratory of Cardiovascular Medicine, Chinese Academy of Medical Science, Peking Union Medical College This protocol presents the establishment of deep hypothermic circulatory arrest in rats, which can be applied to investigate systemic inflammatory response syndrome, ischemia/reperfusion injury, oxidative stress, neuroinflammation, etc. Biochemistry Transplantation of Neonatal Mouse Cardiac Macrophages into Adult Mice Yandong Li1, Jie Feng1, Yan Li1, Jianqiu Pei1, Shengshou Hu1, Yu Nie1,2 1Fuwai Hospital & Chinese Academy of Medical Sciences, State Key Laboratory of Cardiovascular Disease, Cardiovascular Institute, National Center for Cardiovascular Diseases, Peking Union Medical College, 2National Health Commission Key Laboratory of Cardiovascular Regenerative Medicine, Fuwai Central-China Hospital, Central-China Subcenter of National Center for Cardiovascular Diseases We provide a protocol for neonatal cardiac macrophage separation and transplantation into an adult mouse heart, which could be a promising way to promote cardiac repair. Immunology and Infection Functionalization of Atomic Force Microscope Cantilevers with Single-T Cells or Single-Particle for Immunological Single-Cell Force Spectroscopy Jiahuan Chen1, Ying Xu2, Yan Shi2, Tie Xia2 1State Key Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 2Institute for Immunology, School of Medicine, Tsinghua University We present a protocol to functionalize atomic force microscope (AFM) cantilevers with a single T cell and bead particle for immunological studies. Procedures to probe single-pair T cell-dendritic cell binding by AFM and to monitor the real-time cellular response of macrophages to a single solid particle by AFM with fluorescence imaging are shown. Medicine Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo Mingming Liu1, Xiaoyan Zhang1, Bingwei Li1, Bing Wang1, Qingbin Wu1, Fei Shang1, Ailing Li1, Hongwei Li1, Ruijuan Xiu1 1Institute of Microcirculation, Chinese Academy of Medical Sciences & Peking Union Medical College Pancreatic islet microvascular vasomotion regulates islet blood distribution and maintains the physiological function of islet β cells. This protocol describes using a laser Doppler monitor to determine the functional status of pancreatic islet microvascular vasomotion in vivo and to assess the contributions of pancreatic islet microcirculation to pancreatic-related diseases. Developmental Biology A Standardized Method for Measuring Internal Lung Surface Area via Mouse Pneumonectomy and Prosthesis Implantation Zhe Liu1,2, Siling Fu2,3, Nan Tang2 1Department of Life Sciences, Peking University, 2National Institute of Biological Sciences, Beijing, 3Graduate School of Peking Union Medical College Internal lung surface area (ISA) is a critical criterion for assessing lung morphology and physiology in lung diseases and injury-induced alveolar regeneration. We describe here a standardized method that can minimize the measurement bias for ISA in both lung pneumonectomy and prosthesis implantation mouse models. Developmental Biology Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors Wei Wen1, Jian-Ping Zhang1, Wanqiu Chen2, Cameron Arakaki2, Xiaolan Li1,2, David Baylink2, Gary D. Botimer3, Jing Xu1, Weiping Yuan1, Tao Cheng1,4,5,6,7, Xiao-Bing Zhang1,2 1State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, 2Division of Regenerative Medicine, Department of Medicine, Loma Linda University, 3Department of Orthopaedic Surgery, Loma Linda University, 4Center for Stem Cell Medicine, Chinese Academy of Medical Sciences, 5Department of Stem Cell & Regenerative Medicine, Peking Union Medical College, 6Collaborative Innovation Center for Cancer Medicine, 7Tianjin Key Laboratory of Blood Cell Therapy and Technology This protocol describes a detailed method for efficient generation of integration-free iPSCs from human adult peripheral blood cells. With the use of four oriP/EBNA-based episomal vectors to express the reprogramming factors, KLF4, MYC, BCL-XL, or OCT4 and SOX2, thousands of iPSC colonies can be obtained from 1 mL of peripheral blood.