RIKEN Center for Biosystems Dynamics Research 6 articles published in JoVE Biology Isolation Method for Long-Term and Short-Term Hematopoietic Stem Cells Katsuyuki Nishi1,2, Akiomi Nagasaka1,2, Taro Sakamaki1,2, Kay Sadaoka1,2, Masanori Miyanishi1,2 1Hematopoietic Stem Cell Biology and Medical Innovation (HSCBMI), Department of Pediatrics, Kobe University Graduate School of Medicine, 2RIKEN Center for Biosystems Dynamics Research We present a step-by-step protocol for the isolation of long-term hematopoietic stem cells (LT-HSCs) and short-term HSCs (ST-HSCs) using the Hoxb5 reporter system. Bioengineering Preparation of Mesh-Shaped Engineered Cardiac Tissues Derived from Human iPS Cells for In Vivo Myocardial Repair Takeichiro Nakane1,2,6, Mosha Abulaiti1,2, Yuko Sasaki1, William J. Kowalski3, Bradley B. Keller4,5,7, Hidetoshi Masumoto1,2 1Clinical Translational Research Program, RIKEN Center for Biosystems Dynamics Research, 2Department of Cardiovascular Surgery, Graduate School of Medicine, Kyoto University, 3Laboratory of Stem Cell and Neuro-Vascular Biology, Cell and Developmental Biology Center, National Institutes of Health, 4Kosair Charities Pediatric Heart Research Program, Cardiovascular Innovation Institute, University of Louisville, 5Department of Pediatrics, School of Medicine, University of Louisville, 6Department of Cardiovascular Surgery, Mitsubishi Kyoto Hospital, 7 The present protocol generates mesh-shaped engineered cardiac tissues containing cardiovascular cells derived from human induced pluripotent stem cells to allow the investigation of cell implantation therapy for heart diseases. Medicine A Rabbit Venous Interposition Model Mimicking Revascularization Surgery using Vein Grafts to Assess Intimal Hyperplasia under Arterial Blood Pressure Hiroomi Nishio1,3, Kenji Minatoya1, Hidetoshi Masumoto1,2 1Department of Cardiovascular Surgery, Graduate School of Medicine, Kyoto University, 2Clinical Translational Research Program, RIKEN Center for Biosystems Dynamics Research, 3Department of Cardiovascular Surgery, Takamatsu Red Cross Hospital The present protocol aims to experimentally create venous intimal hyperplasia by subjecting veins to arterial blood pressure for developing strategies to attenuate venous intimal hyperplasia following revascularization surgery using vein grafts. Biochemistry An Integrated Raman Spectroscopy and Mass Spectrometry Platform to Study Single-Cell Drug Uptake, Metabolism, and Effects Ahmed Ali1,2, Yasmine Abouleila1,2, Arno Germond1 1Biodynamics Research Center (BDR), RIKEN, 2Research Center, Misr International University This protocol presents an integrated Raman spectroscopy-mass spectrometry (MS) platform that is capable of achieving single-cell resolution. Raman spectroscopy can be used to study cellular response to drugs, while MS can be used for targeted and quantitative analysis of drug uptake and metabolism. Biochemistry Proteome-wide Quantification of Labeling Homogeneity at the Single Molecule Level Simon Leclerc1,2, Youri Arntz2, Yuichi Taniguchi1,3 1Laboratory for Cell Systems Control, RIKEN Center for Biosystems Dynamics Research, RIKEN, 2Laboratoire de Biomatériaux et Bioimagerie, INSERM, 3PRESTO, Japan Science and Technology Agency Here, we present a protocol to assess the labeling homogeneity for each protein species in a complex protein sample at the single molecule level. Biology TChIP-Seq: Cell-Type-Specific Epigenome Profiling Mari Mito1,2, Mitsutaka Kadota3, Shinichi Nakagawa1,4, Shintaro Iwasaki2,5 1RNA Biology Laboratory, RIKEN, 2RNA Systems Biochemistry Laboratory, RIKEN Cluster for Pioneering Research, 3Laboratory for Phyloinformatics, RIKEN Center for Biosystems Dynamics Research, 4RNA Biology Laboratory, Faculty of Pharmaceutical Sciences, Hokkaido University, 5Department of Computational Biology and Medical Sciences, Graduate School of Frontier Sciences, University of Tokyo We describe a step-by-step protocol for tandem chromatin immunoprecipitation sequencing (tChIP-Seq) that enables the analysis of cell-type-specific genome-wide histone modification.
