National Health Research Institutes 7 articles published in JoVE Biology Induction of Petite Colonies in Candida glabrate via Rose Bengal-Mediated Photodynamic Therapy Cai-Ying Yang*1,2, Jia-Horung Hung*3, Chi-Jung Wu4,5, Zhao-Xiang Wang1,2, Shih-Han Wang6, Hao-Chun Liaw2, I-Huang Lin3, Chun-Keung Yu1,6, Tak-Wah Wong2,7,8 1Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, 2Department of Dermatology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, 3Department of Ophthalmology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, 4National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, 5Department of Internal Medicine, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, 6Insititute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, 7Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, 8Center of Applied Nanomedicine, National Cheng Kung University The significance of petite colonies in Candida spp. drug resistance has not been fully explored. Antimicrobial photodynamic therapy (aPDT) offers a promising strategy against drug-resistant fungal infections. This study demonstrates that rose bengal-mediated aPDT effectively deactivates Candida glabrata and induces petite colonies, presenting a unique procedure. Bioengineering Establishing Single-Cell Based Co-Cultures in a Deterministic Manner with a Microfluidic Chip Cheng-Kun He1,2, Ya-Wen Chen3, Ssu-Han Wang3, Chia-Hsien Hsu1,2 1Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, 2Ph.D. Program in Tissue Engineering and Regenerative Medicine, National Chung Hsing University, 3National Institute of Cancer Research, National Health Research Institutes This report describes a microfluidic chip-based method to set up a single cell culture experiment in which high-efficiency pairing and microscopic analysis of multiple single cells can be achieved. Biology In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity Wan-Shan Yang1, Mel Campbell2, Hsing-Jien Kung2,3,4,5, Pei-Ching Chang1,6 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. Bioengineering An Ultrasonic Tool for Nerve Conduction Block in Diabetic Rat Models Yee Fun Lee1, Chou-Ching Lin2, Jung-Sung Cheng1, Gin-Shin Chen1 1Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, 2Department of Neurology, National Cheng Kung University Hospital This work presents the methodology of applying high intensity-focused ultrasound to block the action potentials of diabetic neuropathic nerves. Developmental Biology Transfer of Mammary Gland-forming Ability Between Mammary Basal Epithelial Cells and Mammary Luminal Cells via Extracellular Vesicles/Exosomes Meng-Chieh Lin*1, Shih-Yin Chen*1, Pei-Lin He1, Wen-Ting Luo1, Hua-Jung Li1 1Institute of Cellular and System Medicine, National Health Research Institutes This protocol describes methods for purifying, quantitating, and characterizing extracellular vesicles (EVs)/exosomes from non-adherent/mesenchymal mammary epithelial cells and for using them to transfer mammary gland-forming ability to luminal mammary epithelial cells. EVs/exosomes derived from stem-like mammary epithelial cells can transfer this cell property to cells that ingest the EVs/exosomes. Bioengineering A Microfluidic Platform for High-throughput Single-cell Isolation and Culture Ching-Hui Lin1,2, Hao-Chen Chang1,2, Chia-Hsien Hsu1,2,3 1Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Taiwan, 2Tissue Engineering and Regenerative Medicine, National Chung Hsing University, 3Institute of NanoEngineering and MicroSystems, National Tsing Hua University Here, we present a protocol for isolating and culturing single cells with a microfluidic platform, which utilizes a new microwell design concept to allow for high-efficiency single cell isolation and long-term clonal culture. Immunology and Infection Assessment of the Immunomodulatory Properties of Human Mesenchymal Stem Cells (MSCs) Pei-Ju Hsu1, Ko-Jiunn Liu2, Ying-Yin Chao1, Huey-Kang Sytwu3, B. Linju Yen1 1Regenerative Medicine Research Group, Institute of Cellular & System Medicine, National Health Research Institutes (NHRI), 2National Institute of Cancer Research, National Health Research Institutes (NHRI), 3Institute of Microbiology & Immunology, National Defense Medical Center The immunomodulatory properties of human mesenchymal stem cells (MSC) appear increasingly relevant for clinical application. Using a co-culture system of MSCs and peripheral blood leukocytes pre-stained with the fluorescent dye carboxyfluorescein succinimidyl ester (CFSE), we describe the in vitro assessment of MSC immunomodulation on effector leukocyte proliferation and specific subpopulations.
