Flow Cytometry Protocol for Cell Death Analysis in Glioblastoma Organoids: A Technical Note v1
Year: 2025
Type: preprint
Abstract: Here, we present a protocol to quantify cell death in glioblastoma organoids (GBOs) using flow cytometry. Single-cell suspensions are generated through combined enzymatic and mechanical dissociation, followed by permeabilization and nuclear staining with propidium iodide or Hoechst 33258 to detect DNA fragmentation. Following treatment with temozolomide (TMZ) and lomustine (CCNU) for up to 288 hours, flow cytometric analysis revealed cell death rates of up to 63%, with CCNU demonstrating greater efficacy than TMZ. The extent of cell death varied between GBO populations but was consistent across replicates derived from the same GBO population. The protocol is scalable, cost-effective, and suitable for high-throughput applications in translational cancer research.
Authors: Anna-Laura Potthoff, Meng-Chun Hsieh, Ahmad Melhem, Susanna S. Ng, Barbara E. F. Pregler, Annika Vieregge, Markus Raspe, Lea L. Friker, Thomas Zeyen, Julian P. Layer, Andreas Dolf, Marieta I. Toma, Andreas Waha, Torsten Pietsch, Mike-Andrew Westhoff, Hartmut Vatter, Michael Hölzel, Ulrich Herrlinger, Matthias Schneider
Institutions: University Hospital Bonn, University of Bonn
Language: English
Subfield: Genetics
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W7084033351
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