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Erasmus University Medical Center

7 ARTICLES PUBLISHED IN JoVE

Biology

Isolation of Early Hematopoietic Stem Cells from Murine Yolk Sac and AGM

Kelly Morgan¹, Michael Kharas¹, Elaine Dzierzak², D. Gary Gilliland^1,3

¹Department of Hematology and Oncology, Brigham and Women's Hospital and Harvard Medical School, ²Department of Cell Biology and Genetics, Erasmus University Medical Center, ³Department of Medicine, Howard Hughes Medical Institute, Brigham and Women's Hospital and Harvard Medical School

This video shows how to micro-dissect the yolk sac and aorta-gonad-mesonephros region from embryos and use flow cytometry to sort hematopoietic stem cells.

Cancer Research

Four-color Fluorescence Immunohistochemistry of T-cell Subpopulations in Archival Formalin-fixed, Paraffin-embedded Human Oropharyngeal Squamous Cell Carcinoma Samples

Simone Punt¹, Robert J. Baatenburg de Jong², Ekaterina S. Jordanova^1,3

¹Department of Pathology, Leiden University Medical Center, ²Department of Otorhinolaryngology and Head and Neck Surgery, Erasmus University Medical Center, ³Center for Gynecological Oncology Amsterdam, VU Medical Center

Multiparameter fluorescence immunohistochemistry can be used to assess the number, relative distribution, and localization of immune cell populations in the tumor microenvironment. This manuscript describes the use of this technique to analyze T-cell subpopulations in oropharyngeal cancer.

JoVE Journal

The Organoid Reconstitution Assay (ORA) for the Functional Analysis of Intestinal Stem and Niche Cells

Matthias Schewe¹, Andrea Sacchetti¹, Mark Schmitt¹, Riccardo Fodde¹

¹Department of Pathology, Erasmus MC Cancer Institute, Erasmus University Medical Center

Intestinal organoid cultures are established from whole crypts and do not allow the analysis of self-renewal and differentiation in a cell-specific fashion. This protocol describes reconstitution of sorted stem (Lgr5⁺) and niche (Paneth) cells, which give rise to organoids while enabling their prior biochemical and genetic modification and functional analysis.

Cancer Research

Immunoglobulin Gene Sequence Analysis In Chronic Lymphocytic Leukemia: From Patient Material To Sequence Interpretation

Andreas Agathangelidis^*1, Lesley Ann Sutton^*2,3, Anastasia Hadzidimitriou¹, Cristina Tresoldi⁴, Anton W. Langerak⁵, Chrysoula Belessi⁶, Frederic Davi⁷, Richard Rosenquist^2,3, Kostas Stamatopoulos^1,2, Paolo Ghia⁸

¹Institute of Applied Biosciences, Centre for Research and Technology Hellas, ²Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, ³Department of Molecular Medicine and Surgery, Karolinska Institutet, ⁴Division of Immunology, Transplantation and Infectious, IRCCS San Raffaele Scientific Institute, ⁵Department of Immunology, Laboratory for Medical Immunology, Erasmus University Medical Center, ⁶Hematology Department, Nikea General Hospital, ⁷Assistance publique - Hôpitaux de Paris (AP-HP), Hopital Pitié-Salpêtrière, Department of Hematology, and UPMC University Paris 06, UMRS 1138, ⁸Division of Experimental Oncology, IRCCS Istituto Scientifico San Raffaele and Università Vita-Salute San Raffaele

Herein, we present a protocol that details the technical aspects and essential requirements to ensure robust IG gene sequence analysis in patients with chronic lymphocytic leukemia (CLL), based on the accumulated experience of the European Research initiative on CLL (ERIC).

Biochemistry

Detection of Protein Ubiquitination Sites by Peptide Enrichment and Mass Spectrometry

Karel Bezstarosti¹, Lennart van der Wal¹, Jeroen A. A. Demmers¹

¹Proteomics Center, Erasmus University Medical Center

We present a method for the purification, detection, and identification of diGly peptides that originate from ubiquitinated proteins from complex biological samples. The presented method is reproducible, robust, and outperforms published methods with respect to the level of depth of the ubiquitinome analysis.

Genetics

Validating Whole Genome Nanopore Sequencing, using Usutu Virus as an Example

Bas B. Oude Munnink¹, David F. Nieuwenhuijse¹, Reina S. Sikkema¹, Marion Koopmans¹

¹ErasmusMC, Department of Viroscience, WHO Collaborating Centre for Arbovirus and Viral Hemorrhagic Fever Reference and Research, Erasmus University Medical Center

We previously validated a protocol for amplicon-based whole genome Usutu virus (USUV) sequencing on a nanopore sequencing platform. Here, we describe the methods used in more detail and determine the error rate of the nanopore R10 flow cell.

Medicine

Intraoperative Assessment of Resection Margins in Oral Cavity Cancer: This is the Way

Yassine Aaboubout^1,2, Elisa M. Barroso^1,3,4, Mahesh Algoe¹, Patricia C. Ewing-Graham¹, Ivo ten Hove^3,6, Hetty Mast³, José A. Hardillo², Aniel Sewnaik², Dominiek A. Monserez², Stijn Keereweer², Brend P. Jonker³, Cornelia G. F. van Lanschot², Roeland W. H. Smits², Maria R. Nunes Soares^1,4, Lars Ottevanger¹, Sanne E. Matlung¹, Paul A. Seegers⁵, Vera van Dis¹, Robert M. Verdijk¹, Eppo B. Wolvius³, Peter J. Caspers⁴, Tom C. Bakker Schut⁴, Robert J. Baatenburg de Jong², Gerwin J. Puppels⁴, Senada Koljenović¹

¹Department of Pathology, Erasmus MC University Medical Center, ²Department of Otorhinolaryngology and Head and Neck Surgery, Erasmus MC University Medical Center, ³Department of Oral and Maxillofacial Surgery, Erasmus MC University Medical Center, ⁴Department of Dermatology, Erasmus MC University Medical Center, ⁵PALGA foundation, The nationwide network and registry of histo- and cytopathology, ⁶Department of Oral and Maxillofacial Surgery, Leiden University Medical Center

The goal of this protocol is to provide a clear overview of specimen-driven intraoperative assessment of resection margins. It is encouraged to implement this protocol to improve patient care at other institutes.