Tongji University 11 articles published in JoVE Biology Visualizing Mitophagy with Fluorescent Dyes for Mitochondria and Lysosome Bilin Liu1,2, Anqi Li1, Yuan Qin3, Lei Chen1, Meng Gao1, Guohua Gong1,2 1Institute for Regenerative Medicine, Shanghai East Hospital, School of Life Sciences and Technology, Tongji University, 2Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, 3Department of Pharmacy, Shanghai East Hospital, Tongji University Mitophagy is the primary mechanism of mitochondrial quality control. However, the evaluation of mitophagy in vivo is hindered by the lack of reliable quantitative assays. Presented here is a protocol for the observation of mitophagy in living cells using a cell-permeant green-fluorescent mitochondria dye and a red-fluorescent lysosome dye. Biology Induction and Diverse Assessment Indicators of Experimental Autoimmune Encephalomyelitis Chun Wang*1, Jie Lv*1, Wei Zhuang1,2, Ling Xie1, Guangyu Liu1, Kaidireya Saimaier1, Sanxing Han1, Changjie Shi1, Qiuhong Hua1, Ru Zhang1, Guangfeng Shi3, Changsheng Du1 1Putuo People's Hospital, Shanghai Key Laboratory of Signaling and Disease Research, School of Life Sciences and Technology, Tongji University, 2National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 3Department of Infectious Diseases, Shanghai Key Laboratory of Infectious Diseases and Biosafety Emergency Response, National Medical Center for Infectious Diseases, Huashan Hospital, Fudan University The present protocol describes the induction of experimental autoimmune encephalomyelitis in a mouse model using myelin oligodendrocyte glycoprotein and monitoring the disease process using a clinical scoring system. Experimental autoimmune encephalomyelitis-related symptoms are analyzed using mouse femur micro-computed tomography analysis and open field test to assess the disease process comprehensively. Neuroscience Evaluating Postural Control and Lower-extremity Muscle Activation in Individuals with Chronic Ankle Instability Lulu Yin1, Zhangqi Lai2, Xiaoyue Hu2, Kun Liu3, Lin Wang2 1Department of Critical Care Medicine, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, 2School of Kinesiology, Shanghai University of Sport, 3 Individuals with chronic ankle instability (CAI) exhibit postural control deficiency and delayed muscle activation of lower extremities. Computerized dynamic posturography combined with surface electromyography provides insights into the coordination of the visual, somatosensory, and vestibular systems with muscle activation regulation to maintain postural stability in individuals with CAI. Environment Studying Neurobehavioral Effects of Environmental Pollutants on Zebrafish Larvae Bin Zhang1,2, Xinyue Yang2, Jing Zhao3, Ting Xu2,4, Daqiang Yin2,4 1State Key Laboratory of Marine Geology, Tongji University, 2Key Laboratory of Yangtze River Water Environment, Ministry of Education, College of Environmental Science and Engineering, Tongji University, 3Shanghai Collaborative Innovation Centre for WEEE Recycling, WEEE Research Center of 10 Shanghai Polytechnic University, 4Shanghai Institute of Pollution Control and Ecological Security A detailed experimental protocol is presented in this paper for the evaluation of neurobehavioral toxicity of environmental pollutants using a zebrafish larvae model, including the exposure process and tests for neurobehavioral indicators. Environment A Dual-Functional Electroactive Filter Towards Simultaneously Sb(III) Oxidation and Sequestration Fuqiang Liu1, Fang Li1,2, Chensi Shen1,2, Zhiwei Wang2,3, Wolfgang Sand1,4, Yanbiao Liu1,2 1Textile Pollution Controlling Engineering Center of Ministry of Environmental Protection, College of Environmental Science and Engineering, Donghua University, 2Shanghai Institute of Pollution Control and Ecological Security, 3State Key Laboratory of Pollution Control and Resource Reuse, School of Environmental Science and Engineering, Tongji University, 4Institute of Biosciences, Freiberg University of Mining and Technology A protocol for the rational design of a dual-functional electroactive filter consisting of carbon nanotubes and titanate nanowires is reported and their environmental applications towards Sb(III) oxidation and sequestration is presented. Neuroscience Identifying Cell Surface Markers of Primary Neural Stem and Progenitor Cells by Metabolic Labeling of Sialoglycan Qing-Ran Bai*1, Lu Dong*2, Qin Shen1 1Brain and Spinal Cord Innovative Research Center of Tongji Hospital, School of Life Sciences and Technology, Tongji University and Frontier Science Research Center for Stem Cells of Ministry of Education, 2College of Chemistry and Molecular Engineering, Peking-Tsinghua Center for Life Sciences, Beijing National Laboratory for Molecular Sciences, Synthetic and Functional Biomolecules Center, and Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, Peking University Presented here is a protocol that combines an in vitro neural-endothelial co-culture system and metabolic incorporation of sialoglycan with bioorthogonal functional groups to expand primary neural stem and progenitor cells and label their surface sialoglycoproteins for imaging or mass-spectrometry analysis of cell surface markers. Environment Using Caenorhabditis elegans for Studying Trans- and Multi-Generational Effects of Toxicants Zhuo Li1,2, Fangting Ai3, Jing Zhang4, Zhenyang Yu1,2, Daqiang Yin1,2 1College of Environmental Sciences and Engineering, Tongji University, 2Shanghai Institute of Pollution Control and Ecological Security, 3Jiaxing Tongji Institute of Environment, 4College of Ecological Technique and Engineering, Shanghai Institute of Technology Trans- and multi-generational effects of persistent chemicals are essential in judging their long-term consequences in the environment and on the human health. We provide novel detailed methods for studying trans- and multi-generational effects using free-living nematode Caenorhabditis elegans. Developmental Biology Formation of Human Periodontal Ligament Cell Spheroids on Chitosan Films Xiangzhen Yan*1, Xing Ran*2, Siying Xia1, Yanan Yang1, Min Zhou1, Chunxue Yuan3, Lijun Luo1 1Department of Periodontology, School & Hospital of Stomatology, Tongji University, Shanghai Engineering Research Center of Tooth Restoration and Regeneration, 2Department of Periodontology, Dental Diseases Prevention & Treatment Center of Jiading District, 3College of Materials Science and Engineering, Tongji University Here, we present protocols of culturing human periodontal ligament (PDL) cell spheroids by chitosan films. The culture of three-dimensional (3D) cellular spheroids provides an alternative to conventional tissue culture polystyrene (TCPS) culture system. Environment A 3-dimensional (3D)-printed Template for High Throughput Zebrafish Embryo Arraying Tianyu Yu1,2, Yue Jiang1, Sijie Lin1 1College of Environmental Science and Engineering, State Key Laboratory of Pollution Control and Resource Reuse, Tongji University, 2UNEP-Tongji Institute of Environment for Sustainable Development, Tongji University Here, we present a protocol to design and fabricate a zebrafish embryo arraying template, followed by a detailed procedure on the use of such template for high throughput zebrafish embryo arraying into a 96-well plate. Developmental Biology In Vivo Tracking of Human Adipose-derived Mesenchymal Stem Cells in a Rat Knee Osteoarthritis Model with Fluorescent Lipophilic Membrane Dye Meng Li*1,2, Ming Hao*1,2, Dong Jiang3, Yanxi Chen4, Wen Wang1,2 1Cellular Biomedicine Group, China, 2Cellular Biomedicine Group, California, 3Department of Orthopedics, Shanghai TCM-Integrated Hospital, 4Department of Orthopedics, Shanghai East Hospital, Tongji University This protocol describes an efficient way to monitor the cell persistence and biodistribution of human adipose-derived mesenchymal stem cells (haMSCs) by far-red fluorescence labeling in a rat knee osteoarthritis (KOA) model via intra-articular (IA) injection. Developmental Biology Differentiating Chondrocytes from Peripheral Blood-derived Human Induced Pluripotent Stem Cells Yueying Li2, Yong Hai1, Jiayu Chen3, Tie Liu1 1Department of Orthopedics, Beijing Chao-Yang Hospital, Capital Medical University, 2Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, 3Clinical and Translational Research Center of Shanghai First Maternity & Infant Hospital, School of Life Sciences and Technology, Tongji University We present a protocol to generate a chondrogenic lineage from human peripheral blood (PB) via induced pluripotent stem cells (iPSCs) using an integration-free method, which includes embryoid body (EB) formation, fibroblastic cells expansion, and chondrogenic induction.