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Juntendo University Faculty of Medicine

4 ARTICLES PUBLISHED IN JoVE

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.

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 Okubo⁵, Hiromitsu Komiyama¹, Yutaka Kojima¹, Michitoshi Goto¹, Sonoko Habu³, Okio Hino², Kazuhiro Sakamoto¹, Akira Orimo²

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.

Cancer Research

Genome-Wide Analysis of DNA Methylation in Gastrointestinal Cancer

Kiichi Sugimoto^1,2, Hirotaka Momose², Tomoaki Ito^1,3, Hajime Orita³, Koichi Sato³, Kazuhiro Sakamoto², Malcolm V. Brock¹

¹Department of Surgery, The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, ²Department of Coloproctological Surgery, Juntendo University Faculty of Medicine, ³Department of Surgery, Juntendo University Shizuoka Hospital

Herein, we describe a procedure for genome-wide analysis of DNA methylation in gastrointestinal cancers. The procedure is of relevance to studies that investigate relationships between methylation patterns of genes and factors contributing to carcinogenesis in gastrointestinal cancers.

Cancer Research

A Syngeneic Orthotopic Osteosarcoma Sprague Dawley Rat Model with Amputation to Control Metastasis Rate

Shun Ishiyama^1,2,3,4, Casey Kissel⁵, Xin Guo¹, Alexis Howard⁶, Harumi Saeki⁷, Tomoaki Ito⁸, Polina Sysa-Shah⁹, Hajime Orita¹⁰, Kazuhiro Sakamoto⁴, Kathleen Gabrielson^1,2

¹Department of Molecular and Comparative Pathobiology, The Johns Hopkins University School of Medicine, ²Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, ³Department of Surgery, Johns Hopkins University School of Medicine, ⁴Department of Coloproctological Surgery, Juntendo University Faculty of Medicine, ⁵Program for Comparative Medicine, Gene Therapy Program, Perelman School of Medicine, University of Pennsylvania, ⁶Tuskegee College of Veterinary Medicine, ⁷Department of Human Pathology, Juntendo University Faculty of Medicine, ⁸Department of Surgery, Juntendo University Shizuoka Hospital, Juntendo University School of Medicine, ⁹Department of Urology, The Johns Hopkins School of Medicine, ¹⁰Department of Gastroenterology and Minimally Invasive Surgery, Juntendo University Faculty of Medicine

Here, a syngeneic orthotopic implantation followed by an amputation procedure of the osteosarcoma with spontaneous pulmonary metastasis that can be used for preclinical investigation of metastasis biology and development of novel therapeutics is described.