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13 ARTICLES PUBLISHED IN JoVE

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Developmental Biology

Generation of Induced-pluripotent Stem Cells Using Fibroblast-like Synoviocytes Isolated from Joints of Rheumatoid Arthritis Patients
Yeri Alice Rim 1,3, Narae Park 1, Yoojun Nam 1, Ji Hyeon Ju 2,3
1CiSTEM Laboratory, Convergent Research Consortium for Immunologic Disease, Division of Rheumatology, Seoul St. Mary's Hospital, Republic of Korea, 2Division of Rheumatology, Department of Internal Medicine, Seoul St. Mary's Hospital, Institute of Medical Science, Republic of Korea, 3College of Medicine, The Catholic University of Korea, Republic of Korea

Here we describe a protocol for generating human induced-pluripotent stem cells from patient-derived fibroblast-like synoviocytes, using a lentiviral system without feeder cells.

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JoVE Journal

Procedure for Adaptive Laboratory Evolution of Microorganisms Using a Chemostat
Haeyoung Jeong *1, Sang J. Lee *2, Pil Kim 3
1Super-Bacteria Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), 2Microbiomics and Immunity Research Center, Korea Research Institute of Bioscience and Bioengineering (KRIBB), 3Department of Biotechnology, The Catholic University of Korea

Here, we present a protocol to obtain adaptive laboratory evolution of microorganisms under conditions using chemostat culture. Also, genomic analysis of the evolved strain is discussed.

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Developmental Biology

Induced Pluripotent Stem Cell Generation from Blood Cells Using Sendai Virus and Centrifugation
Yeri Alice Rim 1, Yoojun Nam 1, Ji Hyeon Ju 2
1CiSTEM Laboratory, Convergent Research Consortium for Immunologic Disease, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, 2Division of Rheumatology, Department of Internal Medicine, Seoul St. Marys Hospital, Institute of Medical Science, College of Medicine, The Catholic University of Korea

We propose a protocol for reprogramming peripheral blood mononuclear cells (PBMCs) into induced pluripotent stem cells (iPSCs). By plating the transduced blood cells onto matrix-coated plates with centrifugation, iPSCs are successfully induced from floating cells. This technique suggests a simple and effective reprogramming protocol for cells such as PBMCs and CBMCs.

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Developmental Biology

Chondrogenic Differentiation Induction of Adipose-derived Stem Cells by Centrifugal Gravity
Yeonsue Jang 1, Hyerin Jung 1, Ji Hyeon Ju 1
1CiSTEM Laboratory, Convergent Research Consortium for Immunologic Disease, Division of Rheumatology, Seoul St. Mary’s Hospital, College of Medicine, The Catholic University of Korea

Mechanical stress can induce the chondrogenic differentiation of stem cells, providing a potential therapeutic approach for the repair of impaired cartilage. We present a protocol to induce the chondrogenic differentiation of adipose-derived stem cells (ASCs) using centrifugal gravity (CG). CG-induced upregulation of SOX9 results in the development of chondrogenic phenotypes.

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Developmental Biology

Chondrogenic Pellet Formation from Cord Blood-derived Induced Pluripotent Stem Cells
Yoojun Nam 1, Yeri Alice Rim 1, Ji Hyeon Ju 2
1CiSTEM Laboratory, Convergent Research Consortium for Immunologic Disease, Division of Rheumatology, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, 2Division of Rheumatology, Department of Internal Medicine, Seoul St. Mary's Hospital, Institute of Medical Science, College of Medicine, The Catholic University of Korea

Here, we propose a protocol for chondrogenic differentiation from cord blood mononuclear cell-derived human induced pluripotent stem cells.

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Medicine

The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice
Ji-Eun Kim 1, Kyung-Ok Cho 1
1Department of Pharmacology, Catholic Neuroscience Institute, College of Medicine, The Catholic University of Korea

This manuscript describes a method of inducing status epilepticus by systemic pilocarpine injection and of monitoring spontaneous recurrent seizures in live animals using a wireless telemetry video and electroencephalogram system. This protocol can be utilized for studying the pathophysiologic mechanisms of chronic epilepsy, epileptogenesis, and acute seizures.

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Developmental Biology

Generation of 3D Skin Organoid from Cord Blood-derived Induced Pluripotent Stem Cells
Yena Kim 1,2, Ji Hyeon Ju 1,2,3
1CiSTEM Laboratory, Catholic Induced Pluripotent Stem Cell (iPSC) Research Center, College of Medicine, The Catholic University of Korea, 2Department of Biomedicine & Health Science, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, 3Division of Rheumatology, Department of Internal Medicine, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea

We propose a protocol that shows how to differentiate induced pluripotent stem cell-derived keratinocytes and fibroblasts and generate a 3D skin organoid, using these keratinocytes and fibroblasts. This protocol contains an additional step of generating a humanized mice model. The technique presented here will improve dermatologic research.

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Behavior

Transcranial Direct Current Stimulation for Online Gamers
Sang Hoon Lee *1, Jooyeon Jamie Im *2, Jin Kyoung Oh 2, Eun Kyoung Choi 2, Sujung Yoon 3, Marom Bikson 4, In-Uk Song 5, Hyeonseok Jeong 2, Yong-An Chung 2
1Department of Radiology, Yeouido St. Mary's Hospital, College of Medicine, The Catholic University of Korea, 2Department of Radiology, Incheon St. Mary's Hospital, College of Medicine, The Catholic University of Korea, 3Department of Brain and Cognitive Sciences, Ewha Womans University, 4Department of Biomedical Engineering, The City College of New York, 5Department of Neurology, Incheon St. Mary's Hospital, College of Medicine, The Catholic University of Korea

We present a protocol and a feasibility study for applying transcranial direct current stimulation (tDCS) and neuroimaging assessment in online gamers.

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Behavior

Assessment of Memory Function in Pilocarpine-induced Epileptic Mice
Kwang-Mo Park 1, Ji-Eun Kim 1, In-Young Choi 1, Kyung-Ok Cho 1
1Department of Pharmacology, Department of Biomedicine & Health Sciences, Catholic Neuroscience Institute, Institute of Aging and Metabolic Diseases, College of Medicine, The Catholic University of Korea

This article presents experimental procedures for assessing memory impairments in pilocarpine-induced epileptic mice. This protocol can be used to study the pathophysiologic mechanisms of epilepsy-associated cognitive decline, which is one of the most common comorbidities in epilepsy.

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Medicine

Intramyocardial Transplantation of MSC-Loading Injectable Hydrogels after Myocardial Infarction in a Murine Model
Chan Woo Kim 1,2, Chan Joon Kim 1,3, Eun-Hye Park 1,2, Eunmi Lee 1,2, Eunhwa Seong 1,2, Kiyuk Chang 1,2
1Cardiovascular Research Institute for Intractable Disease, College of Medicine, The Catholic University of Korea, 2Division of Cardiology, Department of Internal Medicine, Seoul St. Mary's Hospital, College of Medicine, The Catholic University of Korea, 3Division of Cardiology, Department of Internal Medicine, Uijeongbu St. Mary's Hospital, College of Medicine, The Catholic University of Korea

Stem cell-based therapy has emerged as an efficient strategy to repair injured cardiac tissues after myocardial infarction. We provide an optimal in vivo application for stem cell transplantation using gelatin hydrogels that are able to be enzymatically cross-linked.

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Engineering

Fabrication of Micro-Patterned Chip with Controlled Thickness for High-Throughput Cryogenic Electron Microscopy
Min-Ho Kang 1,2, Minyoung Lee 3,4, Sungsu Kang 3,4, Jungwon Park 3,4,5,6
1Department of Biomedical-Chemical Engineering, The Catholic University of Korea, 2Department of Biotechnology, The Catholic University of Korea, 3School of Chemical and Biological Engineering, and Institute of Chemical Processes, Seoul National University, 4Center for Nanoparticle Research, Institute of Basic Science (IBS), 5Institute of Engineering Research, College of Engineering, Seoul National University, 6Advanced Institutes of Convergence Technology, Seoul National University

A newly developed micro-patterned chip with graphene oxide windows is fabricated by applying microelectromechanical system techniques, enabling efficient and high-throughput cryogenic electron microscopy imaging of various biomolecules and nanomaterials.

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Engineering

Application of Deep Learning-Based Medical Image Segmentation via Orbital Computed Tomography
Yeon Woong Chung 1,2, Dong Gyun Kang 3, Yong Oh Lee 4, Won-Kyung Cho 1,5
1College of Medicine, The Catholic University of Korea, 2Department of Ophthalmology, St. Vincent’s Hospital, 3Yeoncheon Public Medical Center, 4Department of Industrial and Data Engineering, Hongik University, 5Department of Ophthalmology, Uijeongbu St. Mary’s Hospital

An object segmentation protocol for orbital computed tomography (CT) images is introduced. The methods of labeling the ground truth of orbital structures by using super-resolution, extracting the volume of interest from CT images, and modeling multi-label segmentation using 2D sequential U-Net for orbital CT images are explained for supervised learning.

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Medicine

A Rabbit Aortic Valve Stenosis Model Induced by Direct Balloon Injury
Eunmin Kim *1, Eun-Hye Park *1, Jin-Moo Kim 1, Eunmi Lee 1, Sin-Hee Park 1, Chan Woo Kim 1, Ik Jun Choi 1,2, Min-ho Oak 3, Kiyuk Chang 1,4
1Cardiovascular Research Institute for Intractable Disease, College of Medicine, The Catholic University of Korea, 2Division of Cardiology, Incheon St. Mary’s Hospital, College of Medicine, The Catholic University of Korea, 3College of Pharmacy and Natural Medicine Research Institute, Mokpo National University, 4Division of Cardiology, Seoul St. Mary’s Hospital, College of Medicine, The Catholic University of Korea

An appropriate animal model is needed to understand the pathologic mechanisms underlying aortic valve stenosis (AVS) and to evaluate the efficacy of therapeutic interventions. The present protocol describes a new procedure for developing the AVS rabbit model via a direct balloon injury in vivo.

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