National Yang-Ming University

10 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Organotypic Slice Cultures for Studies of Postnatal Neurogenesis

Adam J. Mosa¹, Sabrina Wang^2,3, Yao Fang Tan¹, J. Martin Wojtowicz¹

¹Department of Physiology, University of Toronto, ²Institute of Anatomy and Cell Biology, School of Medicine, National Yang-Ming University, ³Department of Education and Research, Taipei City Hospital

Here we describe a technique for studying hippocampal postnatal neurogenesis using the organotypic slice culture technique. This method allows for in vitro manipulation of adult neurogenesis and allows for the direct application of pharmacological agents to the cultured hippocampus.

Bioengineering

Electrotaxis Studies of Lung Cancer Cells using a Multichannel Dual-electric-field Microfluidic Chip

Hsien-San Hou¹, Hui-Fang Chang¹, Ji-Yen Cheng^1,2,3,4,5

¹Research Center for Applied Sciences, Academia Sinica, ²Institute of Biophotonics, National Yang-Ming University, ³Biophotonics & Molecular Imaging Research Center (BMIRC), National Yang-Ming University, ⁴Department of Mechanical and Mechatronic Engineering, National Taiwan Ocean University, ⁵Ph.D. Program in Microbial Genomics, National Chung Hsing University

Many microfluidic devices have been developed for use in the study of electrotaxis. Yet, none of these chips allows the efficient study of the simultaneous chemical and electric-field (EF) effects on cells. We developed a polymethylmethacrylate-based device that offers better-controlled coexisting EF and chemical stimulation for use in electrotaxis research.

Bioengineering

Designing Microfluidic Devices for Studying Cellular Responses Under Single or Coexisting Chemical/Electrical/Shear Stress Stimuli

Tzu-Yuan Chou¹, Yung-Shin Sun², Hsien-San Hou³, Shang-Ying Wu¹, Yun Zhu¹, Ji-Yen Cheng³, Kai-Yin Lo¹

¹Department of Agricultural Chemistry, National Taiwan University, ²Department of Physics, Fu-Jen Catholic University, ³Research Center for Applied Sciences, Academia Sinica

Micro-fabricated devices integrated with fluidic components provide an in vitro platform for cell studies mimicking the in vivo micro-environment. We developed polymethylmethacrylate-based microfluidic chips for studying cellular responses under single or coexisting chemical/electrical/shear stress stimuli.

Developmental Biology

Long-term Live Imaging of Drosophila Eye Disc

Chia-Kang Tsao^1,2, Hui-Yu Ku^1,2, Y. Henry Sun^1,2

¹Institute of Genomic Sciences, National Yang-Ming University, ²Institute of Molecular Biology, Academia Sinica

This protocol describes a detailed method for the long-term ex vivo culture and live imaging of a Drosophila imaginal disc. It demonstrates photoreceptor differentiation and ommatidial rotation within the 10 h period of live imaging of the eye disc. The protocol is simple and does not require expensive setup.

Biology

In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity

Wan-Shan Yang¹, Mel Campbell², Hsing-Jien Kung^2,3,4,5, Pei-Ching Chang^1,6

¹Institute of Microbiology and Immunology, National Yang-Ming University, ²UC Davis Cancer Center, University of California, Davis, ³Department of Biochemistry and Molecular Medicine, University of California, Davis, ⁴Institute for Translational Medicine, College of Medical Science and Technology, Taipei Medical University, ⁵Division of Molecular and Genomic Medicine, National Health Research Institutes, ⁶Center for Infectious Disease and Cancer Research, Kaohsiung Medical University

Unlike ubiquitin ligases, few E3 SUMO ligases have been identified. This modified in vitro SUMOylation protocol is able to identify novel SUMO E3 ligases by an in vitro reconstitution assay.

Neuroscience

Stereotaxic Surgery for Genetic Manipulation in Striatal Cells of Neonatal Mouse Brains

Shih-Yun Chen¹, Hsiao-Ying Kuo¹, Fu-Chin Liu^1,2

¹Institute of Neuroscience, National Yang-Ming University, ²Brain Research Center, National Yang-Ming University

We describe a protocol of stereotaxic surgery with a homemade head-fixed device for microinjecting reagents into the striatum of neonatal mouse brains. This technique allows genetic manipulation in neuronal cells of specific regions of neonatal mouse brains.

Immunology and Infection

Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells

Chin-Wen Wei¹, Tyng-An Zhou¹, Ivan L. Dzhagalov¹, Chia-Lin Hsu¹

¹Institute of Microbiology and Immunology, National Yang-Ming University

This article illustrates a powerful method to quantify mitochondria or lysosomes in living cells. The combination of lysosome- or mitochondria-specific dyes with fluorescently conjugated antibodies against surface markers allows the quantification of these organelles in mixed cell populations, like primary cells harvested from tissue samples, by using multicolor flow cytometry.

Bioengineering

Wet-spinning-based Molding Process of Gelatin for Tissue Regeneration

Chia-Yu Wang¹, Dewi Sartika², Ding-Han Wang³, Po-Da Hong¹, Juin-Hong Cherng^2,4,5, Shu-Jen Chang^2,6, Cheng-Che Liu⁷, Yi-Wen Wang⁴, Sheng-Tang Wu⁸

¹Department of Materials Science and Engineering, National Taiwan University of Science and Technology, ²Laboratory of Adult Stem Cell and Tissue Regeneration, National Defense Medical Center, ³School of Dentistry, National Yang-Ming University, ⁴Department and Graduate Institute of Biology and Anatomy, National Defense Medical Center, ⁵Department of Gerontological Health Care, National Taipei University of Nursing and Health Sciences, ⁶Division of Rheumatology/Immunology/Allergy, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, ⁷Department of Physiology and Biophysics, Graduate Institute of Physiology, National Defense Medical Center, ⁸Division of Urology, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center

We developed and describe a protocol based on the wet spinning concept, for the construction of gelatin-based biomaterials used for the application of tissue engineering.

Medicine

Sleeve Gastrectomy in Mice using Surgical Clips

Jih-Hua Wei^1,2,3, Che-Hung Yeh⁴, Wei-Jei Lee⁵, Shing-Jong Lin^3,6,7,8,9, Po-Hsun Huang^3,6,7

¹Cardiovascular Division, Internal Medicine Department, Min-Sheng General Hospital, ²Department of Nutrition and Health Sciences, School of Healthcare Management, Kai-Nan University, ³Institute of Clinical Medicine, National Yang-Ming University, ⁴Food Science and Biotechnology, College of Biotechnology and Bioresources, University of Da-Yeh, ⁵Department of Surgery, Min-Sheng General Hospital, ⁶Department of Critical Care Medicine, Taipei Veterans General Hospital, ⁷Division of Cardiology, Department of Internal Medicine, Taipei Veterans General Hospital, ⁸Division of Cardiology, Heart Center, Cheng-Hsin General Hospital, ⁹Taipei Medical University

The prevalence of diabetes and obesity is continuously increasing worldwide. The mechanisms between diabetes, obesity and their associated mortality and co-morbidities need to be further investigated. Here, we present a protocol for sleeve gastrectomy (SG) in animals as an uncomplicated preclinical model of bariatric surgery.

Bioengineering

Electric-Field-Induced Neural Precursor Cell Differentiation in Microfluidic Devices

Hui-Fang Chang¹, Shih-En Chou¹, Ji-Yen Cheng^1,2,3,4

¹Research Center for Applied Sciences, Academia Sinica, ²Institute of Biophotonics, National Yang Ming Chao Tung University, ³Department of Mechanical and Mechatronic Engineering, National Taiwan Ocean University, ⁴College of Engineering, Chang Gung University

In this study, we present a protocol for the differentiation of neural stem and progenitor cells (NPCs) solely induced by direct current (DC) pulse stimulation in a microfluidic system.