Shandong University View Institution's Website 12 articles published in JoVE Biology Exploring Independent Effects of Follicle-Stimulating Hormone In Vivo in a Mouse Model Yanjing Guo1,2,3, Wei Li2,3, Yinghan Wang2 1The Affiliated Hospital of Qingdao University, 2Department of Anesthesiology, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 3Department of Anesthesiology, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University Follicle-stimulating hormone (FSH) in various extragonadal tissues and organs is associated with the pathogenesis of multiple diseases. The ovariectomized and FSH-treated mouse model (OVF) can be used to explore the extragonadal actions of FSH. Biology Establishment and Genetic Manipulation of Murine Hepatocyte Organoids Jiabei Lian*1,2, Xinyuan Meng*1,3, Xiaohui Zhang1,2, Huili Hu1,2 1Key Laboratory of Experimental Teratology, Ministry of Education, Institute of Molecular Medicine and Genetics, School of Basic Medical Sciences, Stem Cell and Regenerative Medicine Research Center, Cheeloo College of Medicine, Shandong University, 2The Research Center of Stem Cell and Regenerative Medicine, Shandong University Cheeloo Medical College, School of Basic Medical Sciences, Shandong University A long-term ex vitro 3D organoid culture system was established from mouse hepatocytes. These organoids can be passaged and genetically manipulated by lentivirus infection of shRNA/ectopic construction, siRNA transfection and CRISPR-Cas9 engineering. Neuroscience Flow Cytometric Analysis of Multiple Mitochondrial Parameters in Human Induced Pluripotent Stem Cells and Their Neural and Glial Derivatives Kristina Xiao Liang*1,2, Anbin Chen*1,2,3,4, Cecilie Katrin Kristiansen1,2, Laurence A. Bindoff1,2 1Department of Clinical Medicine (K1), University of Bergen, 2Neuro-SysMed, Center of Excellence for Clinical Research in Neurological Diseases, Haukeland University Hospital, 3Department of Neurosurgery, Qilu Hospital and Institute of Brain and Brain-Inspired Science, Cheeloo College of Medicine, Shandong University, 4Shandong Key Laboratory of Brain Function Remodeling This study reports a novel approach to measure multiple mitochondrial functional parameters based on flow cytometry and double staining with two fluorescent reporters or antibodies to detect changes in mitochondrial volume, mitochondrial membrane potential, reactive oxygen species level, mitochondrial respiratory chain composition, and mitochondrial DNA. Biology Y-27632 Enriches the Yield of Human Melanocytes from Adult Skin Tissues Chang Liu1, Shuangshuang Wang1, Man Liu2, Fuxiang Bai1, Zhihong Chen3, Ping Wang4, Jun Mi1,5, Xunwei Wu1 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, 3 This paper reports that the addition of Y-27632 to TIVA medium can significantly increase the yield of melanocytes from adult skin tissues. Medicine Spinal Cord Lateral Hemisection and Asymmetric Behavioral Assessments in Adult Rats Xiao-Jing Lin1,2,3, Shaonan Wen2,6, Ling-Xiao Deng3, Heqiao Dai3, Xiaolong Du3, Chen Chen3,4, Melissa J. Walker3,4, Ting-Bao Zhao1,5, Xiao-Ming Xu3 1Department of Spinal Cord Injury and Repair, Trauma and Orthopedics Institute of Chinese PLA, 960th Hospital, Joint Logistics Support Force of PLA, 2Institute of Military Cognition and Brain Sciences, Academy of Military Medical Sciences, Academy of Military Sciences, 3Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, Department of Neurological Surgery and Goodman Campbell Brain and Spine, Indiana University School of Medicine, 4Program in Medical Neuroscience, Paul and Carole Stark Neurosciences Research Institute, Indiana University School of Medicine, 5Department of Orthopedics, the Third Affiliated Hospital, Shandong University, 6Beijing Computing Center, Beijing Academy of Science and Technology Here we describe surgical procedures to produce a reliable spinal cord lateral hemisection (HX) at the 9th thoracic level in adult rats and neurobehavioral assessments designed for detecting asymmetric deficits after such a unilateral injury. Immunology and Infection In Vivo Augmentation of Gut-Homing Regulatory T Cell Induction Hongzheng Bi1,2, Samiksha Wasnik1, David J. Baylink1, Chenfan Liu1,3,4, Xiaolei Tang1,3 1Division of Regenerative Medicine, Department of Medicine, Loma Linda University, 2Zhengzhou University, 3Department of Veterinary Biomedical Sciences, College of Veterinary Medicine, Long Island University, 4Jinan Infectious Disease Hospital, Shandong University Here we present a protocol for in vivo augmentation of gut-homing regulatory T cell induction. In this protocol, dendritic cells are engineered to locally produce high concentrations of the active vitamin D (1,25-dihydroxyvitamin D or 1,25[OH]2D) and the active vitamin A (retinoic acid or RA) de novo. Immunology and Infection High-Efficiency Generation of Antigen-Specific Primary Mouse Cytotoxic T Cells for Functional Testing in an Autoimmune Diabetes Model Howard W. Davidson1, Joseph Ray Cepeda2, Nitin S. Sekhar2, Junying Han2, Ling Gao3, Tomasz Sosinowski1, Li Zhang2 1Barbara Davis Center for Childhood Diabetes, University of Colorado Denver, 2Department of Medicine, Endocrinology, Diabetes & Metabolism, Baylor College of Medicine, 3Scientific Center, Shandong Provincial Hospital affiliated to Shandong University This article describes a protocol for the generation of antigen-specific CD8 T cells, and their expansion in vitro, with the aim of yielding high numbers of functional T cells for use in vitro and in vivo. Environment A Uniaxial Compression Experiment with CO2-Bearing Coal Using a Visualized and Constant-Volume Gas-Solid Coupling Test System Weitao Hou1, Hanpeng Wang1, Wei Wang1, Zhongzhong Liu1, Qingchuan Li1 1Research Center of Geotechnical and Structural Engineering, Shandong University This protocol demonstrates how to prepare a briquette sample and conduct a uniaxial compression experiment with a briquette in different CO2 pressures using a visualized and constant-volume gas-solid coupling test system. It also aims to investigate changes in terms of coal’s physical and mechanical properties induced by CO2 adsorption. Neuroscience Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice Xiaojing Lin1,2, Tingbao Zhao3, Wenhui Xiong1, Shaonan Wen4, Xiaoming Jin1, Xiao-Ming Xu1 1Spinal Cord and Brain Injury Research Group, Stark Neurosciences Research Institute, Department of Neurological Surgery and Goodman and Campbell Brain and Spine, Department of Anatomy and Cell Biology, Indiana University School of Medicine, 2Department of Spinal Cord Injury and Repair, Trauma and Orthopedics Institute of Chinese PLA, General Hospital of Jinan Military Region, 3Department of Orthopedics, Shandong Cancer Hospital, Shandong University, 4Department of Neurobiology, Institute of Basic Medical Sciences, Academy Military Medical Sciences We describe an experimental procedure for measuring neuronal activity through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy in vivo. Developmental Biology A Simplified and Efficient Method to Isolate Primary Human Keratinocytes from Adult Skin Tissue Zhenan Liu1, Jie Wen1,2, Xue Leng1, Qian Zhou1, Changkuo Zhou3, Huaqiang Zhao1, Xunwei Wu1,2 1Shandong Provincial Key Laboratory of Oral Tissue Regeneration and Laboratory for Tissue Engineering and Regeneration, School of Stomatology, Shandong University, 2Suzhou Institute of Shandong University, 3Department of Urology, Qilu Hospital of Shandong University Here we present a protocol to efficiently isolate primary human keratinocytes from adult skin tissues. This method simplifies the conventional procedure by using the ROCK Inhibitor Y-27632 in the inoculation medium to spontaneously separate epidermal cells from dermal cells. Chemistry Elemental-sensitive Detection of the Chemistry in Batteries through Soft X-ray Absorption Spectroscopy and Resonant Inelastic X-ray Scattering Jinpeng Wu1,2, Shawn Sallis2,3, Ruimin Qiao2, Qinghao Li2,4, Zengqing Zhuo2,5, Kehua Dai2,6, Zixuan Guo2,7, Wanli Yang2 1Geballe Laboratory for Advanced Materials, Stanford University, 2Advanced Light Source, Lawrence Berkeley National Laboratory, 3Department of Materials Science and Engineering, Binghamton University, 4School of Physics, National Key Laboratory of Crystal Materials, Shandong University, 5School of Advanced Materials, Peking University Shenzhen Graduate School, 6School of Metallurgy, Northeastern University, 7Department of Chemical Engineering, University of California-Santa Barbara Here, we present a protocol for typical experiments of soft X-ray absorption spectroscopy (sXAS) and resonant inelastic X-ray scattering (RIXS) with applications in battery material studies. Bioengineering Electric Field-controlled Directed Migration of Neural Progenitor Cells in 2D and 3D Environments Xiaoting Meng*1, Wenfei Li*2,3, Fraser Young1, Runchi Gao3, Laura Chalmers3, Min Zhao3, Bing Song1 1School of Dentistry, Cardiff Institute of Tissue Engineering & Repair, Cardiff University, 2Shandong Qianfoshan Hospital, Shandong University School of Medicine, 3Dermatology and Ophthalmology Research, Institute for Regenerative Cures, University of California at Davis This protocol demonstrates methods used to establish 2D and 3D environments in custom-designed electrotactic chambers, which can track cells in vivo/ex vivo using time-lapse recording at the single cell level, in order to investigate galvanotaxis/electrotaxis and other cellular responses to direct current (DC) electric fields (EFs).