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Shantou University Medical College

13 ARTICLES PUBLISHED IN JoVE

JoVE Journal

Enrichment and Purging of Human Embryonic Stem Cells by Detection of Cell Surface Antigens Using the Monoclonal Antibodies TG30 and GCTM-2

Juan Carlos Polanco¹, Bei Wang¹, Qi Zhou¹, Hun Chy¹, Carmel O'Brien¹, Andrew L. Laslett¹

¹Materials Science and Engineering, CSIRO

We describe the use of the monoclonal antibodies TG30 (CD9) and GCTM-2 for the combined detection of cell surface antigens via fluorescence activated cell sorting (FACS) for the identification and enrichment of live human embryonic stem cells (hESC) using positive selection and also the use of negative selection to purge hESCs from a mixed cell population.

Developmental Biology

A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro

Zhaofeng Jia^1,2, Yujie Liang³, Bin Ma^4,5, Xiao Xu^2,6, Jianyi Xiong², Li Duan², Daping Wang^1,2

¹Guangzhou Medical University, ²Shenzhen Key Laboratory of Tissue Engineering, Shenzhen Laboratory of Digital Orthopeadic Engineering, Department of Orthopedics, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), ³Department of Chemistry, The Chinese University of Hong Kong, ⁴School of Biomedical Engineering, Shanghai Jiao Tong University, ⁵Renji Hospital Clinical Stem Cell Research Center, Shanghai Jiao Tong University School of Medicine, ⁶Shantou University Medical College

We present a method to quantify DNA methylation based on the 5-methylcytosine (5-mC) dot blot. We determined the 5-mC levels during chondrocyte dedifferentiation. This simple technique could be used to quickly determine the chondrocyte phenotype in ACI treatment.

Neuroscience

Localization of Odorant Receptor Genes in Locust Antennae by RNA In Situ Hybridization

Xiao Xu¹, Yinwei You^1,2, Long Zhang¹

¹Department of Entomology, China Agricultural University, ²Bio-tech Research Center, Shandong Academy of Agricultural Sciences

This paper describes a detailed and highly effective RNA in situ hybridization protocol particularly for low-level expressed Odorant Receptor (OR) genes, as well as other genes, in insect antennae using digoxigenin (DIG)-labeled or biotin-labeled probes.

Biology

Magnetic-Activated Cell Sorting Strategies to Isolate and Purify Synovial Fluid-Derived Mesenchymal Stem Cells from a Rabbit Model

Zhaofeng Jia^1,2,3, Yujie Liang^4,5, Xingfu Li^2,3, Xiao Xu^1,2,3, Jianyi Xiong^2,3, Daping Wang^1,2,3, Li Duan^2,3

¹Postgraduate institution, Guangzhou Medical University, ²Guangdong Provincial Research Center for Artificial Intelligence and Digital Orthopedic Technology, ³Shenzhen Key Laboratory of Tissue Engineering, Shenzhen Laboratory of Digital Orthopaedic Engineering, Shenzhen Second People's Hospital (The First Hospital Affiliated to Shenzhen University), ⁴Department of Chemistry, Chinese University of Hong Kong, ⁵Shenzhen Kangning Hospital, Shenzhen Mental Health Center

This article presents a simple and economic protocol for the straightforward isolation and purification of mesenchymal stem cells from New Zealand white rabbit synovial fluid.

Medicine

Preparation, Procedures and Evaluation of Platelet-Rich Plasma Injection in the Treatment of Knee Osteoarthritis

Ziming Chen^*1,2, Zhantao Deng^*1, Yuanchen Ma¹, Junxing Liao¹, Qingtian Li¹, Mengyuan Li¹, Hua Liu¹, Ganghong Chen¹, Chaoming Zeng¹, Qiujian Zheng¹

¹Department of Orthopedics, Guangdong General Hospital (Guangdong Academy of Medical Sciences), ²Shantou University Medical College

Knee osteoarthritis is frequently seen in the orthopedic department. We introduce in detail the entire knee osteoarthritis treatment process with platelet-rich plasma injection, including preparation, procedures, and evaluation.

Cancer Research

Performing Data Mining And Integrative Analysis Of Biomarker in Breast Cancer Using Multiple Publicly Accessible Databases

Min-na Chen¹, De Zeng², Zhuo-qun Zheng³, Zheng Li³, Jian-le Wu³, Jun-yu Jin³, He-jia Wang³, Cui-zhen Huang¹, Hao-yu Lin¹

¹Department of Thyroid and Breast Surgery, First Affiliated Hospital of Shantou University Medical College, ²Department of Medical Oncology, Cancer Hospital of Shantou University Medical College, ³Shantou University Medical College

Here, we present a protocol to explore the biomarker and survival predictor of breast cancer based on the comprehensive analysis of pooled clinical datasets derived from a variety of publicly accessible databases, using the strategy of expression, correlation and survival analysis step by step.

Medicine

Applying a Three-dimensional Uniaxial Mechanical Stimulation Bioreactor System to Induce Tenogenic Differentiation of Tendon-Derived Stem Cells

Ziming Chen^*1, Peilin Chen^*1, Rui Ruan^*1, Lianzhi Chen¹, Jun Yuan¹, David Wood¹, Tao Wang¹, Ming Hao Zheng¹

¹Centre of Orthopaedic Translational Research, Medical School, University of Western Australia

A three-dimensional uniaxial mechanical stimulation bioreactor system is an ideal bioreactor for tenogenic-specific differentiation of tendon-derived stem cells and neo-tendon formation.

Immunology and Infection

Modeling Hepatitis B Virus Infection in Non-Hepatic 293T-NE-3NRs Cells

Xiaoyue Sun^*1,2,3, Xiaoqiang Yang^*1,2,3,4, Chui Zeng^1,2,3, Fayed Attia Koutb Megahed^1,2,3,5, Qi Zhou^1,2,3, Tingdang Liu^1,2,3, Waqas Iqbal^1,2,3, Pingnan Sun^1,2,3, Xiaoling Zhou^1,2,3

¹Stem Cell Research Center, Shantou University Medical College, ²The Center for Reproductive Medicine, Shantou University Medical College, ³Guangdong Provincial Key Laboratory of Infectious Diseases and Molecular Immunopathology, Shantou University Medical College, ⁴Medical Research Center, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, ⁵Department of Nucleic Acid Researches, Genetic Engineering and Biotechnology Research Institute, General Autority - City of Scientific Researches and Technological Applications

This manuscript describes a detailed protocol for Hepatitis B virus (HBV) infection in novel engineered 293T cells (293T-NE-3NRs, expressing human NTCP, HNF4α, RXRα and PPARα) and traditional hepatic cells (HepG2-NE, expressing human NTCP).

Biology

Large-Scale Preparation of Synovial Fluid Mesenchymal Stem Cell-Derived Exosomes by 3D Bioreactor Culture

Li Duan^1,2, Xingfu Li^1,2, Xiao Xu^1,2, Limei Xu^1,2, Daping Wang^1,2, Kan Ouyang^1,2, Yujie Liang^1,3

¹Department of Orthopedics, The First affiliated hospital of Shenzhen University, Shenzhen Second People's Hospital, ²Guangdong Provincial Research Center for Artificial Intelligence and Digital Orthopedic Technology, Shenzhen Second People's Hospital, ³Department of Child and Adolescent Psychiatry, Shenzhen Kangning Hospital, Shenzhen Mental Health Center, Shenzhen Key Laboratory for Psychological Healthcare & Shenzhen Institute of Mental Health

Here, we present a protocol to produce a large number of GMP-grade exosomes from synovial fluid mesenchymal stem cells using a 3D bioreactor.

Immunology and Infection

An Efficient Method for Directed Hepatocyte-Like Cell Induction from Human Embryonic Stem Cells

Qi Zhou^*1,2,3, Xiaoling Xie^*1,2,3, Zhiqian Zhong^1,2,3, Pingnan Sun^1,2,3, Xiaoling Zhou^1,2,3

This manuscript describes a detailed protocol for differentiation of human embryonic stem cells (hESCs) into functional hepatocyte-like cells (HLCs) by continuously supplementing Activin A and CHIR99021 during hESC differentiation into definitive endoderm (DE).

Medicine

A Two-Step Method for Percutaneous Transhepatic Choledochoscopic Lithotomy

Zeming Chen¹, Zhuo Hua¹, Runzhui Lin¹, Hongcai Zhuang¹, Xingmu Liu¹

¹Department of General Surgery, Second Affiliated Hospital, Shantou University Medical College

This "two-step method" significantly improved the success rate of percutaneous transhepatic choledochoscopy and achieved a better prognosis of intrahepatic and extrahepatic choledocholithiasis.

Medicine

Application of Optical Coherence Tomography to a Mouse Model of Retinopathy

Xiaoting Mai^*1, Shaofen Huang^*1, Wen Chen¹, Tsz Kin Ng^1,2,3, Haoyu Chen¹

¹Joint Shantou International Eye Center, Shantou University and the Chinese University of Hong Kong, ²Shantou University Medical College, ³Department of Ophthalmology and Visual Sciences, the Chinese University of Hong Kong

Here, we describe an in vivo imaging technique using optical coherence tomography to facilitate the diagnosis and quantitative measurement of retinopathy in mice.

Engineering

Triplet Fusion Upconversion Nanocapsule Synthesis

Tracy H. Schloemer^1,2, Samuel N. Sanders¹, Qi Zhou², Pournima Narayanan³, Manchen Hu², Mahesh K. Gangishetty¹, Daniel Anderson¹, Michael Seitz^1,2, Arynn O. Gallegos², R. Christopher Stokes¹, Daniel N. Congreve^1,2

¹Rowland Institute, Harvard University, ²Department of Electrical Engineering, Stanford University, ³Department of Chemistry, Stanford University

This protocol details the synthesis of upconversion nanocapsules for subsequent use in photopolymerizable resins for triplet fusion upconversion-facilitated volumetric 3D printing.