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Friedrich-Alexander University of Erlangen-Nuremberg

4 ARTICLES PUBLISHED IN JoVE

Medicine

Isolation of Human Islets from Partially Pancreatectomized Patients

Gregor Bötticher¹, Dorothèe Sturm¹, Florian Ehehalt¹, Klaus P. Knoch², Stephan Kersting¹, Robert Grützmann¹, Gustavo B. Baretton³, Michele Solimena², Hans D. Saeger¹

¹Department of GI-, Thorax- and Vascular Surgery, University Hospital Carl Gustav Carus, University of Technology Dresden, ²Molecular Diabetology, Paul Langerhans Institute Dresden, ³Department of Pathology, University Hospital Carl Gustav Carus, University of Technology Dresden

The supply of type 2 diabetic islets for research is insufficient. Here we share our protocol for isolating islets from patients undergoing partial pancreatectomy. This approach represents a unique venue for obtaining islets from type 2 diabetic and clinically matched non-diabetic subjects in adequate numbers for basic and clinical studies.

Medicine

Improved Protocol For Laser Microdissection Of Human Pancreatic Islets From Surgical Specimens

Dorothée Sturm^1,2, Lorella Marselli³, Florian Ehehalt^1,2, Daniela Richter¹, Marius Distler², Stephan Kersting^1,2, Robert Grützmann², Krister Bokvist⁴, Philippe Froguel⁵, Robin Liechti⁶, Anne Jörns⁷, Paolo Meda⁸, Gustavo Bruno Baretton⁹, Hans-Detlev Saeger², Anke M. Schulte¹⁰, Piero Marchetti³, Michele Solimena¹

¹Molecular Diabetology, Paul Langerhans Institute Dresden, ²Department of GI-, Thoracic- and Vascular Surgery, University Hospital Carl Gustav Carus, University of Technology Dresden, ³Department of Endocrinology and Metabolism, Metabolic Unit University of Pisa, ⁴Labs DC0522, Lilly Corporate Center, ⁵Genomics, Faculty of Medicine Imperial College London, ⁶Vital-IT, SIB Swiss Institute of Bioinformatics, ⁷Clinical Biochemistry, Hannover Medical School, ⁸Cell Physiology and Metabolism, Medical School, University of Geneva, ⁹Department of Pathology, University Hospital Carl Gustav Carus, University of Technology Dresden, ¹⁰R&D DIAB Division / Translational Medicine, Sanofi-Aventis

Laser microdissection is a technique that allows the recovery of selected cells from minute amounts of parenchyma. Here we describe a protocol for acquiring human pancreatic islets from surgical specimens to be used for transcriptomic studies. Our protocol improves the intrinsic autofluorescence of human beta cells, thus facilitating their collection.

Cancer Research

Isolation of Human Endothelial Cells from Normal Colon and Colorectal Carcinoma - An Improved Protocol

Elisabeth Naschberger¹, Daniela Regensburger¹, Clara Tenkerian¹, Melanie Langheinrich², Felix B. Engel³, Carol Geppert⁴, Arndt Hartmann⁴, Robert Grützmann², Vera S. Schellerer^*2, Michael Stürzl^*1

¹Division of Molecular and Experimental Surgery, Department of Surgery, University Medical Center Erlangen, Friedrich-Alexander University of Erlangen-Nuremberg, ²Department of Surgery, University Medical Center Erlangen, Friedrich-Alexander University of Erlangen-Nuremberg, ³Division of Nephropathology, Department of Pathology, University Medical Center Erlangen, Friedrich-Alexander University of Erlangen-Nuremberg, ⁴Department of Pathology, University Medical Center Erlangen, Friedrich-Alexander University of Erlangen-Nuremberg

Tumor endothelial cells are important determinants of the tumor microenvironment and the course of the disease. Here, a protocol for the isolation of pure and viable endothelial cells from human colorectal carcinoma and normal colon to be used in drug testing and pathogenesis research is described.

Medicine

Investigating Intestinal Barrier Breakdown in Living Organoids

Marco Bardenbacher¹, Barbara Ruder², Natalie Britzen-Laurent¹, Elisabeth Naschberger¹, Christoph Becker², Ralph Palmisano³, Michael Stürzl^*1, Philipp Tripal^*1,3

¹Division of Molecular and Experimental Surgery, Department of Surgery, Translational Research Center, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg and Universitätsklinikum Erlangen, ²Department of Medicine 1, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg and Universitätsklinikum Erlangen, ³Optical Imaging Centre Erlangen (OICE), Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg

Here we describe a technique to quantify the barrier integrity of small intestinal organoids. The fact that the method is based on living organoids enables the sequential investigation of different barrier integrity modulating substances or combinations thereof in a time-resolved manner.