Princess Maxima Center for Pediatric Oncology 6 articles published in JoVE Biology Generation of Human Kidney Tubuloids from Tissue and Urine Camilla Calandrini1,2, Jarno Drost1,2 1Princess Máxima Center for Pediatric Oncology, 2Oncode Institute Human kidney tubuloid cultures represent a valuable in vitro model to study kidney physiology and disease. Tubuloids can be established from kidney tissue (healthy and diseased) as well as urine, the latter representing an easily obtainable and less invasive source of research material. Neuroscience A High-Throughput Image-Guided Stereotactic Neuronavigation and Focused Ultrasound System for Blood-Brain Barrier Opening in Rodents Rianne Haumann*1,2, Elvin ’t Hart*2, Marc P. P. Derieppe2, Helena C. Besse3, Gertjan J. L. Kaspers1,2, Eelco Hoving2, Dannis G. van Vuurden1,2, Esther Hulleman1,2, Mario Ries3 1Amsterdam UMC, Vrije Universiteit Amsterdam, Pediatric Oncology, Cancer Center Amsterdam, 2Princess Máxima Center for Pediatric Oncology, 3Imaging Division, Utrecht University The blood-brain barrier (BBB) can be temporarily disrupted with microbubble-mediated focused ultrasound (FUS). Here, we describe a step-by-step protocol for high-throughput BBB opening in vivo using a modular FUS system accessible for non-ultrasound experts. Biology Single-Cell Resolution Three-Dimensional Imaging of Intact Organoids Ravian L. van Ineveld*1,2,3, Hendrikus C.R. Ariese*1,2,3, Ellen J. Wehrens1,2,3, Johanna F. Dekkers*1,2,3,4, Anne C. Rios*1,2,3 1Princess Máxima Center for Pediatric Oncology, 2Department of Cancer Research, Oncode Institute, Hubrecht Institute-KNAW Utrecht, 3Cancer Genomics Center (CGC), 4Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW) and University Medical Center (UMC) Utrecht The entire 3D structure and cellular content of organoids, as well as their phenotypic resemblance to the original tissue can be captured using the single-cell resolution 3D imaging protocol described here. This protocol can be applied to a wide range of organoids varying in origin, size and shape. Genetics Characterizing Mutational Load and Clonal Composition of Human Blood Axel Rosendahl Huber1, Freek Manders1, Rurika Oka1, Ruben van Boxtel1 1Princess Máxima Center for Pediatric Oncology Somatic mutation patterns in cells reflect previous mutagenic exposure and can reveal developmental lineage relationships. Presented here is a methodology to catalogue and analyze somatic mutations in individual hematopoietic stem and progenitor cells. Cancer Research Comprehensive Protocol to Sample and Process Bone Marrow for Measuring Measurable Residual Disease and Leukemic Stem Cells in Acute Myeloid Leukemia Jacqueline Cloos*1,2, Jeffrey R. Harris*3, Jeroen J.W.M. Janssen1, Angele Kelder1, F. Huang3, Gerrit Sijm1, Maike Vonk1, Alexander N. Snel1, Jennifer R. Scheick1, Willemijn J. Scholten1, Jannemieke Carbaat-Ham1, Dennis Veldhuizen1, Diana Hanekamp1, Yvonne J.M. Oussoren-Brockhoff1, Gertjan J.L. Kaspers2,4, Gerrit J. Schuurhuis1, A. Kate Sasser3, Gert Ossenkoppele1 1Department of Hematology, VU University Medical Center, 2Pediatric Oncology/Hematology, VU University Medical Center, 3Janssen Research & Development, LLC, 4Princess Máxima Center for Pediatric Oncology Detection of minimal or measurable residual disease (MRD) is an important prognostic biomarker for refining risk assessment and predicting relapse in acute myeloid leukemia (AML). These comprehensive guidelines and recommendations with best practices for consistent and accurate identification and detection of MRD, may aid in making effective AML treatment decisions. Cancer Research Using RNA-sequencing to Detect Novel Splice Variants Related to Drug Resistance in In Vitro Cancer Models Rocco Sciarrillo1,2,3, Anna Wojtuszkiewicz1, Irsan E. Kooi4, Valentina E. Gómez3, Ugo Boggi5, Gerrit Jansen6, Gert-Jan Kaspers1,7, Jacqueline Cloos*1, Elisa Giovannetti*3,8,9 1Department of Pediatric Oncology/Hematology, VU University Medical Center, 2Department of Hematology, VU University Medical Center, 3Department of Medical Oncology, VU University Medical Center, 4Department of Clinical Genetics, VU University Medical Center, 5Division of General and Transplant Surgery, Azienda Ospedaliera Universitaria Pisana, Universita’ di Pisa, 6Amsterdam Immunology and Rheumatology Center, VU University Medical Center, 7Princess Máxima Center for Pediatric Oncology, 8Cancer Pharmacology Lab, AIRC Start-Up Unit, University of Pisa, 9Institute of Nanoscience and Nanotechnology, CNR-Nano Here we describe a protocol aimed at investigating the impact of aberrant splicing on drug resistance in solid tumors and hematological malignancies. To this goal, we analyzed the transcriptomic profiles of parental and resistant in vitro models through RNA-seq and established a qRT-PCR based method to validate candidate genes.
