JoVE Logo

Sign In

Shanghai Jiaotong University School of Medicine

18 ARTICLES PUBLISHED IN JoVE

image

Developmental Biology

Derivation of Cardiac Progenitor Cells from Embryonic Stem Cells
Ieng Lam Lei 1, Lei Bu 2, Zhong Wang 1
1Cardiac Surgery, University of Michigan, 2Leon H Charney Division of Cardiology, New York University School of Medicine

In this protocol, derivation of cardiac progenitor cells from both mouse and human embryonic stem cells will be illustrated. A major strategy in this protocol is to enrich cardiac progenitor cells with flow cytometry using fluorescent reporters engineered into the embryonic stem cell lines.

image

Developmental Biology

Efficient Generation of hiPSC Neural Lineage Specific Knockin Reporters Using the CRISPR/Cas9 and Cas9 Double Nickase System
Shenglan Li *1,2, Haipeng Xue *1,2, Bo Long 1,2,5, Li Sun 1,2,6, Tai Truong 1,2,4,7, Ying Liu 1,2,3
1Department of Neurosurgery, The University of Texas Health Science Center at Houston, 2Center for Stem Cell and Regenerative Medicine, Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, 3The Senator Lloyd & B. A. Bentsen Center for Stroke Research, Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, 4Summer Research Program, Office of Educational Programs, The University of Texas Health Science Center at Houston, 5Department of Anesthesiology, Shengjing Hospital, China Medical University, 6Department of Oncology, Renji Hospital, Shanghai Jiaotong University School of Medicine, 7Biology Department, University of West Georgia

Genome editing tools such as the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas (CRISPR-associated) system have greatly improved gene targeting efficiency in human induced pluripotent stem cells (hiPSCs). This manuscript describes a protocol for generating lineage specific hiPSC reporter using CRISPR/Cas system assisted homologous recombination.

image

Developmental Biology

Using In Vivo and Tissue and Cell Explant Approaches to Study the Morphogenesis and Pathogenesis of the Embryonic and Perinatal Aorta
Ashish Misra 1, Zhonghui Feng 1, Jiasheng Zhang 1, Zhi-Yin Lou 2,3, Daniel M. Greif 1
1Yale Cardiovascular Research Center, Section of Cardiovascular Medicine, Department of Internal Medicine, Yale University School of Medicine, 2Department of Neurology, Yale University School of Medicine, 3Department of Neurology, Xinhua Hospital, Shanghai Jiaotong University School of Medicine

Protocols for studying the embryonic and perinatal murine aorta using in vivo clonal analysis and fate mapping, aortic explants, and isolated smooth muscle cells are detailed here. These diverse approaches facilitate the investigation of the morphogenesis of the embryonic and perinatal aorta in normal development and the pathogenesis in disease.

image

JoVE Journal

Screening Bioactive Nanoparticles in Phagocytic Immune Cells for Inhibitors of Toll-like Receptor Signaling
Hong Yang 1, Shan Yu Fung 2, Aihua Bao 1, Qiang Li 1, Stuart E. Turvey 2
1Department of Respiratory Medicine, Shanghai First People's Hospital, Shanghai Jiaotong University School of Medicine, 2Department of Pediatrics, BC Children's Hospital and University of British Columbia

Toll-like receptor (TLR) signaling plays an important role in the pathophysiology of many human inflammatory diseases, and regulating TLR responses by bioactive nanoparticles is anticipated to be beneficial in many inflammatory conditions. THP-1 cell-based reporter cells provide a versatile and robust screening platform for identifying novel inhibitors of TLR signaling.

image

Neuroscience

In Vitro Characterization of the Electrophysiological Properties of Colonic Afferent Fibers in Rats
Youqiang Meng 1, Li Dong 1, Biying Sun 1, Ping Luo 1, Guohua Zhang 1, Weifang Rong 1
1Hongqiao International Institute of Medical Research, Tongren Hospital and Department of Physiology, Faculty of Basic Medical Sciences, Shanghai Jiaotong University School of Medicine

Abnormal sensory function underlies visceral pain and other symptoms of functional and inflammatory bowel diseases. A protocol for the electrophysiological recording of the colonic afferent nerves in an ex vivo rat colorectum preparation is presented here.

image

Behavior

Bouncing Ball with a Uniformly Varying Velocity in a Metronome Synchronization Task
Yingyu Huang *1, Li Gu *1, Junkai Yang *1, Xiang Wu 1
1Department of Psychology, Sun Yat-Sen University

The purpose of this protocol is to introduce the application of a bouncing ball with a uniformly varying velocity in a metronome synchronization task.

image

Medicine

A Detailed Protocol for Physiological Parameters Acquisition and Analysis in Neurosurgical Critical Patients
Xiang Wu 1,2, Guoyi Gao 1,2, Junfeng Feng 1,2, Qing Mao 1,2, Jiyao Jiang 1,2
1Department of Neurosurgery, Renji Hospital, Shanghai Jiaotong University School of Medicine, 2Shanghai Institute of Head Trauma

Recently, we have developed a portable multimodality monitoring system for the monitoring of various physiological parameters in neurosurgical critical patients. Detailed protocols on how to use this multimodality monitoring system are presented here.

image

Developmental Biology

A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation
Qinggang Dai *1, Yujiao Han *2, Furong Xie *1, Xuhui Ma 3, Zhan Xu 2, Xiao Liu 1, Weiguo Zou 2, Jun Wang 1
1Department of Pediatric Dentistry, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, National Clinical Research Center of Stomatology, 2State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, University of Chinese Academy of Sciences, 3Department of Oral and Maxillofacial-Head and Neck Oncology, Ninth People's Hospital, Shanghai Jiaotong University School of Medicine

This manuscript describes a protocol to isolate and culture osteoclasts in vitro from mouse bone marrow, and to study the role of the mammalian/mechanistic target of rapamycin complex 1 in osteoclast formation.

image

Medicine

Surgical Treatment for Benign Prostatic Hyperplasia: Holmium Laser Enucleation of the Prostate (HoLEP).
Huan Xu *1, Xiang Wan *1, Meng Gu 1, Yanbo Chen 1, Qiling Shi 1, Qi Chen 1, Zhong Wang 1
1Department of Urology, Shanghai 9th People's Hospital, Shanghai Jiaotong University School of Medicine

Here we present a safe and efficient protocol, holmium laser enucleation of the prostate, to treat benign prostatic hyperplasia.

image

Cancer Research

A GPC3-targeting Bispecific Antibody, GPC3-S-Fab, with Potent Cytotoxicity
Yanlan Wang 1,2, Jiayu Liu 1,2, Haitao Pan 1,2, Jieyu Xing 1,2, Xiaoqiong Wu 1,2, Qing Li 1,2, Zhong Wang 1,2
1School of Pharmaceutical Sciences, Sun Yat-Sen University, 2Center for Cellular & Structural Biology, Sun Yat-Sen University

Here, we present a protocol to produce a bispecific antibody GPC3-S-Fab in Escherichia coli. The purified GPC3-S-Fab has potent cytotoxicity against GPC3 positive liver cancer cells.

image

Chemistry

A New Straightforward Method for Lipophilicity (logP) Measurement using 19F NMR Spectroscopy
Zhong Wang 1, Benjamin F. Jeffries 1, Hannah R. Felstead 1, Neil J. Wells 1, Elisabetta Chiarparin 2, Bruno Linclau 1
1Chemistry, University of Southampton, 2Medicinal Chemistry, Oncology, IMED Biotech Unit, AstraZeneca

A novel and straightforward variation of the shake-flask method was developed for accurate lipophilicity measurement of fluorinated compounds by 19F NMR spectroscopy.

image

Bioengineering

Protein Kinase C-delta Inhibitor Peptide Formulation using Gold Nanoparticles
Hisato Konoeda 1, Hong Yang 2, Chengliang Yang 1, Annette Gower 1, Chun Xu 1, Wei Zhang 1, Mingyao Liu 1,3
1Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, 2Respiratory Medicine Research Laboratory, Institute of Translation Medicine, Shanghai General Hospital, Shanghai Jiaotong University, 3Institute of Medical Science, Faculty of Medicine, University of Toronto

We have previously used a gold nanoparticle peptide hybrid to intravenously deliver a synthetic peptide, protein kinase C-delta inhibitor, which reduced ischemia-reperfusion-induced acute lung injury. Here we show the detailed protocol of the drug formulation. Other intracellular peptides can be formulated similarly.

image

Medicine

Intra-Abdominal Pressure Measurements in Neurocritical Patients
Xuxu Xu *1, Zhenghui He *1, Yingchi Shan 2, Qing Mao 1, Junfeng Feng 1, Guoyi Gao 2
1Department of Neurosurgery, Renji Hospital, Shanghai Jiaotong University School of Medicine, 2Department of Neurosurgery, Shanghai General Hospital, Shanghai Jiaotong University School of Medicine

This protocol details the use of a set of intra-abdominal pressure monitoring systems with advantages of convenience, continuous monitoring, digital visualization, and long-term IAP recording and data storage in neurocritical patients to detect intra-abdominal hypertension, which is used for correlation analysis to guide treatment and predict prognoses.

image

Medicine

In vitro Assessment of Cardiac Reprogramming by Measuring Cardiac Specific Calcium Flux with a GCaMP3 Reporter
Zhaokai Li *1,2, Liu Liu *1, Zhong Wang 1
1Department of Cardiac Surgery, Cardiovascular Center, The University of Michigan, Ann Arbor, 2Department of Cardiovascular Medicine, Xiangya Hospital, Central South University

We describe here, the establishment and application of an Tg(Myh6-cre)1Jmk/J /Gt(ROSA)26Sortm38(CAG-GCaMP3)Hze/J (referred to as αMHC-Cre/Rosa26A-Flox-Stop-Flox-GCaMP3 below) mouse reporter line for cardiac reprogramming assessment. Neonatal cardiac fibroblasts (NCFs) isolated from the mouse strain are converted into induced cardiomyocytes (iCMs), allowing for convenient and efficient evaluation of reprogramming efficiency and functional maturation of iCMs via calcium (Ca2+) flux.

image

Medicine

Interphase Fluorescence in situ Hybridization of Bone Marrow Smears of Multiple Myeloma
Yalan Yu *1, Hui Shen *1, Li Liu 1, Ping Luo 1, Sanyun Wu 1, Jing He 1,2, Xiqin Tong 1, Yufeng Shang 1, Liang Shao 1, Fuling Zhou 1
1Department of Hematology, Zhongnan Hospital of Wuhan University, 2Wuhan National Laboratory for Optoelectronics, Huazhong University of Science and Technology

Here, we present a protocol for improving the success of interphase fluorescence in situ hybridization detection on bone marrow smears from multiple myeloma patients.

image

Medicine

Laparoscopic Radical Antegrade Modular Pancreatosplenectomy via Dorsal-Caudal Artery Approach for Pancreatic Neck-Body Cancer
Zhantao Shen 1,2, Guihao Chen 1,2, Chunbao Zhu 2, Xiang Wu 1,2, Yifeng Liu 1,2, Zhangyuanzhu Liu 1,2, Zhijian Tan 1,2, Xiaosheng Zhong 1,2
1The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, 2The Second Clinical College of Guangzhou University of Chinese Medicine

With advancements in laparoscopic techniques, laparoscopic radical antegrade modular pancreatosplenectomy (L-RAMPS) has been widely recognized. However, owing to several technical difficulties in this procedure, the artery-first approach in L-RAMPS still remains uncommon. Here, we developed the dorsal-caudal artery approach for L-RAMPS, which might be safe and beneficial for pancreatic neck tumors.

image

Medicine

Transcorporal Artificial Urinary Sphincter Cuff Placement in a Case Requiring Revision for Urethral Atrophy
Da-chao Zheng *1, Min-kai Xie *1, Jie-wen Bao 1, Hai-jun Yao 1, Zhong Wang 1
1Department of Urology, Shanghai 9th People’s Hospital, Shanghai Jiaotong University School of Medicine

Here we describe the application of transcorporal artificial urethral sphincter (AUS) placement in a case requiring revision of an artificial urinary sphincter for urethral atrophy.

image

Cancer Research

Ex Vivo Organoid Model of Adenovirus-Cre Mediated Gene Deletions in Mouse Urothelial Cells
Dongbo Xu 1, Li Wang 1, Kyle Wieczorek 1, Yanqing Wang 2, Xiaojing Zhang 2, David W. Goodrich 2, Qiang Li 1,2
1Department of Urology, Roswell Park Comprehensive Cancer Center, 2Department of Pharmacology and Therapeutics, Roswell Park Comprehensive Cancer Center

This protocol describes the process of the generation and characterization of mouse urothelial organoids harboring deletions in genes of interest. The methods include harvesting mouse urothelial cells, ex vivo transduction with adenovirus driving Cre expression with a CMV promoter, and in vitro as well as in vivo characterization.

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved