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Tongji University

25 ARTICLES PUBLISHED IN JoVE

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Biology

Electron Cryotomography of Bacterial Cells
Songye Chen 1, Alasdair McDowall 1,2, Megan J. Dobro 1, Ariane Briegel 1,2, Mark Ladinsky 1,2, Jian Shi 2, Elitza I. Tocheva 1, Morgan Beeby 1,2, Martin Pilhofer 1,2, H. Jane Ding 1, Zhuo Li 1,2, Lu Gan 1, Dylan M. Morris 1, Grant J. Jensen 1,2
1Division of Biology, California Institute of Technology - Caltech, 2Howard Hughes Medical Institute, California Institute of Technology - Caltech

We illustrate here how to use electron cryotomography (ECT) to study the ultrastructure of bacterial cells in near-native states, to "macromolecular" (~4 nm) resolution.

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Biology

Confocal Imaging of Single Mitochondrial Superoxide Flashes in Intact Heart or In Vivo
Guohua Gong 1, Wang Wang 1
1Mitochondria and Metabolism Center, Department of Anesthesiology and Pain Medicine, University of Washington

Confocal scanning microscopy is applied for imaging single mitochondrial events in perfused heart or skeletal muscles in live animal. Real-time monitoring of single mitochondrial processes such as superoxide flashes and membrane potential fluctuations enables the evaluation of mitochondrial function in a physiologically relevant context and during pathological perturbations.

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Developmental Biology

Differentiating Chondrocytes from Peripheral Blood-derived Human Induced Pluripotent Stem Cells
Yueying Li 2, Yong Hai 1, Jiayu Chen 3, Tie Liu 1
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.

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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 Jiang 3, Yanxi Chen 4, Wen Wang 1,2
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.

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Environment

A 3-dimensional (3D)-printed Template for High Throughput Zebrafish Embryo Arraying
Tianyu Yu 1,2, Yue Jiang 1, Sijie Lin 1
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.

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JoVE Journal

A Random-displacement Measurement by Combining a Magnetic Scale and Two Fiber Bragg Gratings
Lianqing Zhu 1,2, Lidan Lu 3,4, Wei Zhuang 2,5, Zhoumo Zeng 3,4, Mingli Dong 1,2
1School of Instrument Science and Opto-electronics Engineering, Beijing Information Science and Technology University, 2Beijing Engineering Research Center of Optoelectronic Information and Instruments, Beijing Key Laboratory for Optoelectronics Measurement Technology, 3School of Precision Instrument & Opto-electronics Engineering, Tianjin University of Science and Technology, 4School of Precision Instrument and Opto-electronics Engineering, State Key Laboratory of Precision Measuring Technology and Instruments, Tianjin University, 5School of Instrument and Opto-electronics Engineering, Hefei University of Technology

A protocol to create a full-range linear displacement sensor, combining two packaged fiber Bragg grating detectors with a magnetic scale, is presented.

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Environment

Using Caenorhabditis elegans for Studying Trans- and Multi-Generational Effects of Toxicants
Zhuo Li 1,2, Fangting Ai 3, Jing Zhang 4, Zhenyang Yu 1,2, Daqiang Yin 1,2
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.

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Cancer Research

A Human Peripheral Blood Mononuclear Cell (PBMC) Engrafted Humanized Xenograft Model for Translational Immuno-oncology (I-O) Research
Zhuo Li *1, Xiao Yang *1, Yilu Zhang 1, Xiaolong Yang 1, Xinxin Cui 1, Yanjuan Zhang 1, Wenfeng Gong 1, Huichen Bai 1, Ning Liu 1, Zhiyu Tang 1, Mingming Guo 1, Kang Li 1, Tong Zhang 1, Lai Wang 1, Xiaomin Song 1
1BeiGene (Beijing) Co., Ltd.

We describe a human peripheral blood mononuclear cell (PBMC) — based humanized xenograft mouse model for translational immuno-oncology research. This protocol could serve as a general guideline for establishing and characterizing similar models for I-O therapy assessment.

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Developmental Biology

Formation of Human Periodontal Ligament Cell Spheroids on Chitosan Films
Xiangzhen Yan *1, Xing Ran *2, Siying Xia 1, Yanan Yang 1, Min Zhou 1, Chunxue Yuan 3, Lijun Luo 1
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.

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Bioengineering

Automated Counterflow Centrifugal System for Small-Scale Cell Processing
Anqi Li 1,2, Stephen Wilson 3, Ian Fitzpatrick 3, Mehri Barabadi 1,2, Siow Teng Chan 1, Mirja Krause 1,2, Gina Diamanta Kusuma 1,2, David James 3, Rebecca Lim 1,2,4
1The Ritchie Centre, Hudson Institute of Medical Research, 2Department of Obstetrics and Gynaecology, Monash University, 3Scinogy, 4Australian Regenerative Medicine Institute, Monash University

Automation is key to upscaling and cost management in cell manufacturing. This manuscript describes the use of a counterflow centrifugal cell processing device for automating the buffer exchange and cell concentration steps for small-scale bioprocessing.

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Environment

A Dual-Functional Electroactive Filter Towards Simultaneously Sb(III) Oxidation and Sequestration
Fuqiang Liu 1, Fang Li 1,2, Chensi Shen 1,2, Zhiwei Wang 2,3, Wolfgang Sand 1,4, Yanbiao Liu 1,2
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.

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Environment

Studying Neurobehavioral Effects of Environmental Pollutants on Zebrafish Larvae
Bin Zhang 1,2, Xinyue Yang 2, Jing Zhao 3, Ting Xu 2,4, Daqiang Yin 2,4
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.

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Neuroscience

Elimination of Serotonergic Neurons by Stereotaxic Injection of 5,7-Dihydroxytryptamine in the Dorsal Raphe Nuclei of Mice
Lining Cao 1, Zhiqiang Zhang 1, Xinyu Lu 1, Guanhao Wang 1, Du Meng 1, Chang Liu 1, Ji Yun 1, Ting Xu 1, Chun Zhao 2, Jianfeng Lu 1
1Institute for Regenerative Medicine, Shanghai East Hospital, Frontier Science Center for Stem Cell Research, School of Life Sciences and Technology, Tongji University, 2Department of Ophthalmology, Shanghai Tenth People's Hospital Affiliated to Tongji University School of Medicine

This protocol describes a dorsal raphe nucleus (DRN)-lesioned mouse model (>90% survival rate in experimental mice) with stable loss of dorsal raphe serotonergic neurons by stereotaxic injection of 5,7-dihydroxytryptamine into the DRN using an angled approach to prevent injury to the superior sagittal sinus.

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Neuroscience

Intra-Arterial Delivery of Neural Stem Cells to the Rat and Mouse Brain: Application to Cerebral Ischemia
Bei Zhang 1, Binoy Joseph 2, Kathryn E. Saatman 2, Lei Chen 2
1College of Public Health, Shaanxi University of Chinese Medicine, 2Spinal Cord and Brain Injury Research Center, Department of Physiology, University of Kentucky

A method for delivering neural stem cells, adaptable for injecting solutions or suspensions, through the common carotid artery (mouse) or external carotid artery (rat) after ischemic stroke is reported. Injected cells are distributed broadly throughout the brain parenchyma and can be detected up to 30 d after delivery.

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Biology

Y-27632 Enriches the Yield of Human Melanocytes from Adult Skin Tissues
Chang Liu 1, Shuangshuang Wang 1, Man Liu 2, Fuxiang Bai 1, Zhihong Chen 3, Ping Wang 4, Jun Mi 1,5, Xunwei Wu 1
1Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, 2Shijiazhuang Shimen Experimental School, 3Qilu Children's Hospital of Shandong University, 4Qilu Hospital of Shandong University, 5Shenzhen Research Institute of Shandong University

This paper reports that the addition of Y-27632 to TIVA medium can significantly increase the yield of melanocytes from adult skin tissues.

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Neuroscience

Evaluating Postural Control and Lower-extremity Muscle Activation in Individuals with Chronic Ankle Instability
Lulu Yin 1, Zhangqi Lai 2, Xiaoyue Hu 2, Kun Liu 3, Lin Wang 2
1Department of Critical Care Medicine, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, 2School of Kinesiology, Shanghai University of Sport, 3Department of Rehabilitation Medicine, Shanghai Jiao Tong University Affiliated Sixth People's Hospital

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.

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Behavior

Individualized rTMS Treatment for Depression using an fMRI-Based Targeting Method
Xin Luo 1, Yiru Hu 1, Runhua Wang 1, Min Zhang 1, Xiaomei Zhong 1, Bin Zhang 1
1Affiliated Brain Hospital of Guangzhou Medical University

The present protocol describes the application of repetitive transcranial magnetic stimulation (rTMS), where a subregion of the dorsolateral prefrontal cortex (DLPFC) with the strongest functional anticorrelation with the subgenual anterior cingulate cortex (sgACC) was located as the stimulation target under the assistance of a fMRI-based neuronavigation system.

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Bioengineering

Isolation and Culture of Primary Human Gingival Epithelial Cells using Y-27632
Zhiwei Xie 1,2,3, Jizhou Shi 4, Min Zong 3, Qiuping Xu 1, Chang Liu 1, Jie Wen 1, Qun Zhang 1, Panpan Liu 1, Guanyi Liu 1, Jing Guo 2,5, Xunwei Wu 1
1Department of Tissue Engineering and Regeneration, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, 2Department of Orthodontics, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, 3Department of Stomatology, Shengli Oilfield Central Hospital, 4Department of Pediatric Surgery, Shengli Oilfield Central Hospital, 5Ningbo Stomatology Hospital of Savaid Stomatology School, Hangzhou Medical College

Here we present a modified method for the isolation and culture of human gingival epithelial cells by adding the Rock inhibitor, Y-27632, to the traditional method. This method is easier, less time-consuming, enhances stem cell properties, and produces larger numbers of high-potential epithelial cells both for the laboratory and for clinical applications.

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Bioengineering

Open-source Toolkit: Benchtop Carbon Fiber Microelectrode Array for Nerve Recording
Julianna M. Richie 1, Paras R. Patel 1, Elissa J. Welle 1, Tianshu Dong 2, Lei Chen 3, Albert J. Shih 2, Cynthia A. Chestek 1,4,5,6
1Department of Biomedical Engineering, University of Michigan, Ann Arbor, 2Department of Mechanical Engineering, University of Michigan, Ann Arbor, 3Department of Mechanical Engineering, University of Massachusetts Lowell, 4Department of Electrical Engineering and Computer Science, University of Michigan, Ann Arbor, 5Neuroscience Graduate Program, University of Michigan, Ann Arbor, 6Robotics Graduate Program, University of Michigan, Ann Arbor

Here, we describe fabrication methodology for customizable carbon fiber electrode arrays for recording in vivo in nerve and brain.

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Biology

Surgically Induced Cardiac Volume Overload by Aortic Regurgitation in Mouse
Jian Wu *1, Shijun Wang *1, Le Kang *1, Chunjie Yang *1, Jianguo Jia 1, Xingxu Wang 2, Ge Wei 1, Xuan Li 1, Jieyun You 2, Yunzeng Zou 1
1Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital and Institutes of Biomedical Sciences, Fudan University, 2Department of Cardiovascular Medicine, Shanghai East Hospital, School of Medicine, Tongji University

This protocol presents a practical guide on the surgery for creation of aortic regurgitation (AR) in the mouse. Assessment of the AR mouse by echocardiography and invasive hemodynamic measurement recapitulates its clinically relevant characteristics of volume overload-induced eccentric hypertrophy, suggesting its promising application in the study of cardiac hypertrophy.

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Biology

Induction and Diverse Assessment Indicators of Experimental Autoimmune Encephalomyelitis
Chun Wang *1, Jie Lv *1, Wei Zhuang 1,2, Ling Xie 1, Guangyu Liu 1, Kaidireya Saimaier 1, Sanxing Han 1, Changjie Shi 1, Qiuhong Hua 1, Ru Zhang 1, Guangfeng Shi 3, Changsheng Du 1
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.

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Bioengineering

A "Plug-And-Display" Nanoparticle Vaccine Platform Based on Outer Membrane Vesicles Displaying SARS-CoV-2 Receptor-Binding Domain
Rang Feng 1, Guo-Cheng Li 1, Hai-Ming Jing 1, Chang Liu 1, Ruo-Yi Xue 1, Quan-Ming Zou 1, Hai-Bo Li 1
1National Engineering Research Center of Immunological Products, Department of Microbiology and Biochemical Pharmacy, College of Pharmacy, Third Military Medical University

The present protocol describes the bioengineering of outer membrane vesicles to be a "Plug-and-Display" vaccine platform, including production, purification, bioconjugation, and characterization.

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Biology

Visualizing Mitophagy with Fluorescent Dyes for Mitochondria and Lysosome
Bilin Liu 1,2, Anqi Li 1, Yuan Qin 3, Lei Chen 1, Meng Gao 1, Guohua Gong 1,2
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.

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Biochemistry

Fluorescent Lateral Flow Immunoassay Based on Quantum Dots Nanobeads
Lingzhi Fan *1, Yue Luo *2, Wannian Yan 1, Huanxing Han 3, Pengfei Zhang 1
1Department of Central Laboratory, Shanghai Skin Disease Hospital, School of Medicine, Tongji University, 2Department of Dermatology, Shanghai Skin Disease Hospital, Institute of Dermatology, School of Medicine, Tongji University, 3Department of Pharmacy, Changzheng Hospital, Naval Medical University

Here, we describe a protocol for the preparation of quantum dot nanobeads (QDNB) and the detection of disease biomarkers using QDNB-based lateral flow immunoassay strips. The test results can be qualitatively assessed under UV light illumination and quantitatively measured using a fluorescent strip reader within 15 min.

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Application of a Dual Upper Limb Task-Oriented Robotic System for the Functional Recovery of the Upper Limb in Stroke Patients

Application of a Dual Upper Limb Task-Oriented Robotic System for the Functional Recovery of the Upper Limb in Stroke Patients
Ya Sun 1,2, Yan Li 2, Danni Xu 2, Linghui Chen 2, Jie Shen 2, Dong Xu 3,4, Han Xu 4, Xu Zhang 4, Xudong Gu 1,2, Jianming Fu 2
1Graduate School of Zhejiang Chinese Medical University, 2The Second Hospital of Jiaxing, 3School of Mechanical Engineering, Tongji University, 4Auckland Tongji Rehabilitation Medical Equipment Research Centre, Tongji Zhejiang College

This experimental protocol outlines the use of a dual upper limb task-oriented robotic system for stroke patients with upper limb dysfunction. The findings indicate that this system can significantly improve stroke patients' upper limb function and daily living activities.

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