University Hospital of Cologne View Institution's Website 6 articles published in JoVE Cancer Research Subculture and Cryopreservation of Esophageal Adenocarcinoma Organoids: Pros and Cons for Single Cell Digestion Ningbo Fan*1, Lisa Raatz*1, Seung-Hun Chon1, Alexander Quaas2, Christiane Bruns1, Yue Zhao1 1Department of General, Visceral, Cancer and Transplantation Surgery, University Hospital Cologne, 2Institute of Pathology, University Hospital Cologne This protocol describes the methods of subculture and cryopreservation of esophageal adenocarcinoma organoids with and without single cell digestion to enable researchers to choose appropriate strategies based on their experimental design. Medicine Customizing a Cryolite Glass Prosthetic Eye Alexander C. Rokohl1, Marc Trester2, Joel M. Mor1, Niklas Loreck1, Konrad R. Koch1, Ludwig M. Heindl1,3 1Department of Ophthalmology, University of Cologne, Faculty of Medicine and University Hospital Cologne, 2Trester-Institute for Ocular Prosthetics and Artificial Eyes, 3Center for Integrated Oncology Aachen-Bonn-Cologne-Dusseldorf This manuscript shows each step of customizing a cryolite glass prosthetic eye including some major advantages of the use of cryolite glass for manufacturing an eye prosthesis compared to poly(methyl methacrylate). In addition, this manuscript gives ophthalmologists better insight into the ocularistic care that could improve interprofessional collaboration. Developmental Biology Visualizing the Node and Notochordal Plate In Gastrulating Mouse Embryos Using Scanning Electron Microscopy and Whole Mount Immunofluorescence Cally Xiao1,2,3, Frank Nitsche4, Hisham Bazzi2,3 1Graduate Program in Pharmacology and Experimental Therapeutics, University of Cologne, 2Department of Dermatology and Venereology, University Hospital of Cologne, 3Cologne Cluster of Excellence in Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, 4Department of General Ecology, Institute for Zoology, Biocenter Cologne, University of Cologne The node and notochordal plate are transient signaling organizers in developing mouse embryos that can be visualized using several techniques. Here, we describe in detail how to perform two of the techniques to study their structure and morphogenesis: 1) scanning electron microscopy (SEM); and 2) whole mount immunofluorescence (WMIF). Medicine The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects Lukas Andereggen3,4,5, Volker Neuschmelting1,6, Michael von Gunten7, Hans Rudolf Widmer5, Jukka Takala1, Stephan M. Jakob1, Javier Fandino1,2, Serge Marbacher1,2 1Department of Intensive Care Medicine, University and Bern University Hospital (Inselspital), 2Department of Neurosurgery, Kantonsspital Aarau, 3 The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures — subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation. Immunology and Infection Murine Model of CD40-activation of B cells Tanja M. Liebig1, Anne Fiedler1, Nela Klein-Gonzalez1, Alexander Shimabukuro-Vornhagen1, Michael von Bergwelt-Baildon1 1Laboratory for Tumor and Transplantation Immunology, Department I of Internal Medicine, University Hospital of Cologne In this video, we demonstrate the procedure of CD40-activation and expansion of murine B cells from splenocytes of C57BL/6 mice, which can be used as a model antigen-presenting cell (APC) to study induction of immunity. Biology Generation of Human CD40-activated B cells Tanja M. Liebig1, Anne Fiedler1, Shahram Zoghi1, Alexander Shimabukuro-Vornhagen1, Michael S. von Bergwelt-Baildon1 1Laboratory for Tumor and Transplantation Immunology and Stem Cell Transplantation Program, University Hospital of Cologne, Department I of Internal Medicine In this video we present the ex vivo generation and expansion of human CD40-activated B cells (CD40-B) from peripheral blood mononuclear cells (PBMC) by stimulation with CD40 ligand and interleukin-4.