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Yonsei University College of Medicine

8 ARTICLES PUBLISHED IN JoVE

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Developmental Biology

Visualization and Quantitative Analysis of Embryonic Angiogenesis in Xenopus tropicalis
Jiyeon Ohk 1, Hosung Jung 1,2
1Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, 2Department of Anatomy, Brain Research Institute, Yonsei University College of Medicine

This protocol demonstrates a fluorescence-based method to visualize the vasculature and to quantify its complexity in Xenopus tropicalis. Blood vessels can be imaged minutes after the injection of a fluorescent dye into the beating heart of an embryo after genetic and/or pharmacological manipulations to study cardiovascular development in vivo.

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Neuroscience

Neurobehavioral Assessments in a Mouse Model of Neonatal Hypoxic-ischemic Brain Injury
MinGi Kim 1,2, Ji Hea Yu 1, Jung Hwa Seo 1,2, Yoon-Kyum Shin 1,2, Soohyun Wi 1,2, Ahreum Baek 1,3, Suk-Young Song 1,5, Sung-Rae Cho 1,2,4,5
1Department and Research Institute of Rehabilitation Medicine, Yonsei University College of Medicine, 2Brain Korea 21 PLUS Project for Medical Science, Yonsei University, 3Department of Rehabilitation Medicine, Yonsei University Wonju College of Medicine, 4Rehabilitation Institute of Neuromuscular Disease, Yonsei University College of Medicine, 5Graduate Program of NanoScience and Technology, Yonsei University

We performed unilateral carotid artery occlusion on postnatal day 7-10 CD-1 mouse pups to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of HI brain injury. We studied neurobehavioral functions in these mice compared to non-operated normal mice.

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Bioengineering

A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer
Alessandra Merenda 1,2, Amanda Andersson-Rolf 1,2, Roxana C. Mustata 1, Taibo Li 1, Hyunki Kim 3, Bon-Kyoung Koo 1,2
1Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge, 2Department of Genetics, University of Cambridge, 3Department of Pathology, Yonsei University College of Medicine

This protocol describes the steps for cloning multiple single guide RNAs into one guide RNA concatemer vector, which is of particular use in creating multi-gene knockouts using CRISPR/Cas9 technology. The generation of double knockouts in intestinal organoids is shown as a possible application of this method.

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Neuroscience

Novel Passive Clearing Methods for the Rapid Production of Optical Transparency in Whole CNS Tissue
Jiwon Woo 1,2,3, Eunice Yoojin Lee 4, Hyo-Suk Park 1,3, Jeong Yoon Park 1,3, Yong Eun Cho 1,2,3
1Department of Neurosurgery, Gangnam Severance Hospital, Yonsei University College of Medicine, 2Brain Korea 21 PLUS Project for Medical Science, Yonsei University, 3The Spine and Spinal Cord Institute, Biomedical Center, Gangnam Severance Hospital, Yonsei University College of Medicine, 4Columbia University College of Physicians and Surgeons

Here, we present two novel methodologies, psPACT and mPACT, for achieving maximal optical transparency and subsequent microscopic analysis of tissue vasculature in the intact rodent whole CNS.

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Immunology and Infection

Two-photon Intravital Imaging of Leukocytes During the Immune Response in Lipopolysaccharide-treated Mouse Liver
Sang A Park *1,2, Young Ho Choe *1,2, Sung Hwan Lee 3, Young-Min Hyun 1,2
1Department of Anatomy, Yonsei University College of Medicine, 2Brain Korea 21 Plus Project for Medical Science, Yonsei University College of Medicine, 3Department of Surgery, Yonsei University College of Medicine

We established a novel surgical protocol for two-photon imaging of live mice liver with minimal invasion. With this technique, we identified the detailed structure of the liver during lipopolysaccharide-induced endotoxemia. We anticipate that this method may be utilized to determine the effectiveness of various reagents treatment to hepatic leukocyte migration.

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Bioengineering

Use of Micro X-ray Computed Tomography with Phosphotungstic Acid Preparation to Visualize Human Fibromuscular Tissue
Jehoon O 1, Hyun-Jin Kwon 1, Shin Hyung Kim 2, Tae-Hyeon Cho 1, Hun-Mu Yang 1,3
1Department of Anatomy, Yonsei University College of Medicine, 2Department of Anesthesiology and Pain Medicine, Anesthesia and Pain Research Institute, Yonsei University College of Medicine, 3Surgical Anatomy Education Centre at the Yonsei University College of Medicine

Micro X-ray computed tomography is effective in obtaining three-dimensional information from undamaged human specimens but has limited success in observing soft tissues. The use of phosphotungstic acid contrast agent can resolve this issue. We implemented this contrast agent to examine human delicate fibromuscular tissues (the orbicularis retaining ligament).

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Immunology and Infection

In vivo Function of Differential Subsets of Cutaneous Dendritic Cells to Induce Th17 Immunity in Intradermal Candida albicans Infection
Jeyun Park 1,2, Sung Hee Kim 1, Joohee Lee 1, Lihua Che 1, Won Seok Roh 1, Chang Ook Park 1,2, Min-Geol Lee 1,3, Tae-Gyun Kim 1,3
1Department of Dermatology, Severance Hospital, Cutaneous Biology Research Institute, Yonsei University College of Medicine, 2Brain Korea 21 FOUR Project for Medical Science, Yonsei University College of Medicine, 3Institute for Immunology and Immunological Diseases, Yonsei University College of Medicine

Here, we demonstrate the in vivo function of cutaneous dendritic cell subsets in Th17 immunity of deep dermal Candida albicans infection.

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Medicine

The Combination of Mechanically Isolated Stromal Vascular Fraction and Fibrin Hydrogel: A Processing Protocol
Pablo Pfister *1, Gregory Reid *2, Tabea Breckwoldt 2, Mauro Vasella 2, Ann-Kathrin Seitz 2, Seung Yong Song 3, Bong-Sung Kim 2
1Department of Surgery, Triemli City Hospital Zurich, 2Department of Plastic Surgery and Hand Surgery, University Hospital Zurich, University of Zurich, 3Institute for Human Tissue Restoration, Department of Plastic and Reconstructive Surgery, Yonsei University College of Medicine

Mechanically isolated stromal vascular fraction (SVF) in combination with a fibrin hydrogel offers an easy and efficient carrier for viable adipose-derived stromal cells for various indications, including tissue engineering and or wound healing purposes. Here, we present the preparation of a mechanical SVF (mSVF)-fibrin hydrogel construct for translational research and clinical application.

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