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Juntendo University

9 ARTICLES PUBLISHED IN JoVE

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Medicine

Experimental Generation of Carcinoma-Associated Fibroblasts (CAFs) from Human Mammary Fibroblasts
Urszula M. Polanska *1, Ahmet Acar *1, Akira Orimo 1,2
1CR-UK Stromal-Tumour Interaction Group, Paterson Institute for Cancer Research, University of Manchester, 2Atopy Research Center, Juntendo University

Carcinoma-associated fibroblasts (CAFs) rich in myofibroblasts present within the tumour stroma, play a major role in driving tumour progression. We developed a coimplantation tumour xengraft model for experimentally generating CAFs from human mammary fibroblasts. The protocol describes how to establish CAF myofibroblasts that acquire an ability to promote tumourigenesis.

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Cancer Research

High-sensitivity Detection of Micrometastases Generated by GFP Lentivirus-transduced Organoids Cultured from a Patient-derived Colon Tumor
Yu Okazawa 1,2, Kosuke Mizukoshi 1,2, Yu Koyama 2,4, Shoki Okubo 5, Hiromitsu Komiyama 1, Yutaka Kojima 1, Michitoshi Goto 1, Sonoko Habu 3, Okio Hino 2, Kazuhiro Sakamoto 1, Akira Orimo 2
1Department of Coloproctological Surgery, Juntendo University Faculty of Medicine, 2Department of Molecular Pathogenesis, Juntendo University Faculty of Medicine, 3Atopy Research Center, Juntendo University Faculty of Medicine, 4Department of Oral Pathobiological Science and Surgery, Tokyo Dental College, 5Department of Gastroenterology, Juntendo University Faculty of Medicine

To allow highly sensitive detection of the disseminating human colorectal cancer (CRC) cells colonizing tissues, we herein show a protocol for efficient transduction of green fluorescent protein (GFP) lentiviral particles into PDX-derived CRC organoid cells prior to their injection into recipient mice, with stereo-fluorescence microscopic observation.

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Medicine

Lung Fixation under Constant Pressure for Evaluation of Emphysema in Mice
Keiko Karasutani 1,2, Hario Baskoro 1, Tadashi Sato 1, Naoko Arano 1, Yohei Suzuki 1, Aki Mitsui 1, Naoko Shimada 1, Yuzo Kodama 1, Kuniaki Seyama 1, Yoshinosuke Fukuchi 1, Kazuhisa Takahashi 1
1Department of Respiratory Medicine, Juntendo University Graduate School of Medicine, Juntendo University, 2Pharmaceutical Planning Group, Otsuka Pharmaceutical Co., Ltd.

Presented here is a useful protocol for lung fixation that creates a stable condition for histological evaluate of lung specimens from a mouse model of emphysema. The main advantage of this model is that it can fix many lungs with the same constant pressure without lung collapse or deflation.

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Neuroscience

Cheek Injection Model for Simultaneous Measurement of Pain and Itch-related Behaviors
Yu Yamanoi 1,2,3,4, Hiroki Kittaka 1,2,5, Makoto Tominaga 1,2,3,6
1Thermal Biology Group, Exploratory Research Center on Life and Living Systems (ExCELLS), 2Division of Cell Signaling, National Institute for Physiological Sciences, 3Department of Physiological Sciences, Graduate University for Advanced Studies (SOKENDAI), 4Research Laboratory, Ikedamohando Co., Ltd., 5Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University, 6Institute for Environmental and Gender-Specific Medicine, Juntendo University

Typically, the mouse neck injection model is used for evaluate pruritogen-induced scratch behaviors. However, the model provides information only on itch, not pain. Here, a cheek injection model is introduced in mice which can be used to simultaneously measure pain and itch-related behaviors.

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Biology

Application of Passive Head Motion to Generate Defined Accelerations at the Heads of Rodents
Takahiro Maekawa *1, Naoyoshi Sakitani *1, Youngjae Ryu 1, Atsushi Takashima 2, Shuhei Murase 1, Julius Fink 3, Motoshi Nagao 1, Toru Ogata 4, Masahiro Shinohara 1, Yasuhiro Sawada 5
1Department of Rehabilitation for Motor Functions, National Rehabilitation Center for Persons with Disabilities, 2Department of Assistive Technology, National Rehabilitation Center for Persons with Disabilities, 3Department of Metabolism and Endocrinology, Graduate School of Medicine, Juntendo University, 4Department of Rehabilitation Medicine, Graduate School of Medicine, The University of Tokyo, 5Department of Clinical Research, National Rehabilitation Center for Persons with Disabilities

The present protocol describes a custom-designed ''passive head motion'' system, which reproduces mechanical accelerations at rodents' heads generated during their treadmill running at moderate velocities. It allows dissecting mechanical factors/elements from the beneficial effects of physical exercise.

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Neuroscience

A Tissue Clearing Method for Neuronal Imaging from Mesoscopic to Microscopic Scales
Kenta Yamauchi 1,2, Shinichiro Okamoto 1,2,3, Megumu Takahashi 1,2,4,5, Masato Koike 2,3, Takahiro Furuta 6, Hiroyuki Hioki 1,2,7
1Department of Neuroanatomy, Juntendo University Graduate School of Medicine, 2Department of Cell Biology and Neuroscience, Juntendo University Graduate School of Medicine, 3Advanced Research Institute for Health Sciences, Juntendo University, 4Department of Neuroscience, Graduate School of Medicine, Kyoto University, 5Japan Society for the Promotion of Science, 6Department of Oral Anatomy and Neurobiology, Graduate School of Dentistry, Osaka University, 7Department of Multi-Scale Brain Structure Imaging, Juntendo University Graduate School of Medicine

The protocol provides a detailed method of neuronal imaging in brain slice using a tissue clearing method, ScaleSF. The protocol includes brain tissue preparation, tissue clarification, handling of cleared slices and confocal laser scanning microscopy imaging of neuronal structures from mesoscopic to microscopic levels.

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Behavior

A Common Marmoset Model of Mother-Infant Intervention for Breastfeeding Disorders in the Presence of Paternal Inhibition and Maternal Neglect
Mayuko Yoda 1,2, Yoshimasa Kamei 3, Hayato Sakurai 4, Hiroko Kakei 4, Ting Tao 1,2,4, Hideo Yamanouchi 4, Tetsuya Kunikata 4, Masanori Hariyama 5, Ricki Colman 6, Mamiko Koshiba 1,4,5
1Graduate School of Sciences and Technology for Innovation, Yamaguchi University, 2Faculty of Health Sciences and Nursing, Juntendo University, 3Obstetrician, Medical Department, Saitama Medical University, 4Department of Pediatrics, Saitama Medical University, 5Graduate School of Information Sciences, Tohoku University, 6Department of Cell and Regenerative Biology and Wisconsin National Primate Research Center, University of Wisconsin, Madison

Here, we present a nonhuman primate model of mother-infant intervention for breastfeeding disorders in the presence of paternal inhibition and maternal neglect. The mate model video complements education to support primate and human caregivers with respect to infants with breastfeeding problems such as pain.

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Immunology and Infection

Adjuvant Activity of Mycobacterium paratuberculosis in Enhancing the Immunogenicity of Autoantigens During Experimental Autoimmune Encephalomyelitis
Davide Cossu 1,2, Yuji Tomizawa 1, Eiichi Momotani 1, Kazumasa Yokoyama 1, Nobutaka Hattori 1,3
1Department of Neurology, Juntendo University, 2Biomedical Research Core Facilities, Juntendo University, 3Neurodegenerative Disorders Collaborative laboratory, RIKEN Center for Brain Science

Here, we present an alternative protocol to actively induce experimental autoimmune encephalomyelitis in C57BL/6 mice, using the immunogenic epitope myelin oligodendrocyte glycoprotein (MOG)35-55 suspended in incomplete Freund's adjuvant containing the heat-killed Mycobacterium avium subspecies paratuberculosis.

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Cancer Research

Facilitating Repeat Intracarotid Injections in Mouse Models by a Novel Injection Site Repair Technique
Daniel Ledbetter 1, Joy Gumin 1, Lynette Phillips 1, Chibawanye Ene 1, Yuzaburo Shimizu 1,2, Frederick F. Lang 1
1Department of Neurosurgery, The University of Texas MD Anderson Cancer Center, 2Department of Neurosurgery, Juntendo University

Repair of the intracarotid artery in a mouse model after injection returns blood flow to the artery without negatively impacting the distribution of the injected material. Injection site repair facilitates subsequent injections through the same artery and prevents cerebral ischemia in mouse strains that lack a complete Circle of Willis.

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