Yong Loo Lin School of Medicine 4 articles published in JoVE Biology Isolation of Primary Rat Hepatocytes with Multiparameter Perfusion Control Inn Chuan Ng*1, Li Zhang*2, Narelle Nichola Yi Ying Shen3, Yun Ting Soong4, Chan Way Ng5, Phoebe Kang Sheing Koh1, Yan Zhou3, Hanry Yu1,3,4,5,6 1Department of Physiology & The Institute for Digital Medicine (WisDM), National University of Singapore, 2College of Agriculture and Biology, Zhongkai University of Agriculture and Engineering, 3Mechanobiology Institute, National University of Singapore, 4Institute of Bioengineering and Nanotechnology, A*STAR, 5NUS Graduate School for Integrative Sciences and Engineering, 6CAMP, Singapore-MIT Alliance for Research and Technology This protocol details the use of a special intravenous catheter, standardized sterile disposable tubing, temperature control complemented by real-time monitoring, and an alarm system for two-step collagenase perfusion procedure to improve the consistency in the viability, yield, and functionality of isolated primary rat hepatocytes. Immunology and Infection In Vitro Differentiation of Mouse Granulocyte-macrophage-colony-stimulating Factor (GM-CSF)-producing T Helper (THGM) Cells Yi Lu1,2,3, Xin-Yuan Fu3, Yongliang Zhang1,2 1Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, 2Immunology Programme, Life Sciences Institute, National University of Singapore, 3Cancer Science Institute of Singapore, Yong Loo Lin School of Medicine, National University of Singapore Here, we present a protocol to differentiate murine granulocyte-macrophage-colony-stimulating-factor-producing T helper (THGM) cells from naive CD4+ T cells, including isolation of naive CD4+ T cells, differentiation of THGM, and analysis of differentiated THGM cells. This method can be applied to studies of the regulation and function of THGM cells. Immunology and Infection A Non-invasive Way to Isolate and Phenotype Cells from the Conjunctiva Tanima Bose1, Aihua Hou2, Ryan Lee2, Louis Tong2,3,4,5, K. George Chandy1 1Lee Kong Chian School of Medicine, Nanyang Technological University, 2Singapore Eye Research Institute, 3Singapore National Eye Center, 4Duke-NUS Medical School, 5Yong Loo Lin School of Medicine The exposed normal ocular surface consists of cornea and conjunctiva. Epithelial cells, goblet cells and immune cells are present in the conjunctiva. Here, a non-invasive, technique of impression cytology is described using an impression cytology device and flow cytometry to analyze immune cells in the conjunctiva. Immunology and Infection Anti-virulent Disruption of Pathogenic Biofilms using Engineered Quorum-quenching Lactonases Song Buck Tay1,3, Jeng Yeong Chow2, Maybelle Kho Go1,3, Wen Shan Yew1,3 1Department of Biochemistry, Yong Loo Lin School of Medicine, 2Department of Chemistry, University of Utah, 3Synthetic Biology Research Consortium, National University of Singapore Quorum-quenching enzymes are anti-virulent and anti-bacterial options that can mitigate pathogenesis without risk of incurring resistance, by preventing the expression of virulence factors and genes associated with antibiotic resistance and biofilm formation. In this study, we report a method that demonstrates the efficacy of quorum-quenching enzymes in bacterial biofilm disruption.
