Optimized culturing yields high success rates and preserves molecular heterogeneity, enabling personalized screening for high-grade gliomas

  • NPJ Precis Oncol. 2025 May 27;9(1):156. doi: 10.1038/s41698-025-00946-1.
Cassandra Posthoorn-Verheul  1 ,  Federica Fabro  1  2  3 ,  Ioannis Ntafoulis  1 ,  Chelsea den Hollander  1 ,  Iris S C Verploegh  1  2 ,  Rutger Balvers  1 ,  Trisha V Kers  1 ,  Jessica Hoogeveen  1 ,  Judith van der Burg  1 ,  Bert Eussen  4 ,  Annelies de Klein  4 ,  Kate J Feller  5 ,  Miao-Ping Chien  5 ,  Clemens M F Dirven  1 ,  Sieger Leenstra  1 ,  Martine L M Lamfers  6
Affiliations
  • 1. Department of Neurosurgery, Brain Tumor Center, Erasmus Medical Center Cancer Institute, Rotterdam, The Netherlands.
  • 2. Department of Cell Biology, Erasmus Medical Center, Rotterdam, The Netherlands.
  • 3. Department of Developmental Biology, Erasmus Medical Center, Rotterdam, The Netherlands.
  • 4. Department of Clinical Genetics, Erasmus Medical Center, Rotterdam, The Netherlands.
  • 5. Department of Molecular Genetics, Erasmus Medical Center, Rotterdam, The Netherlands.
  • 6. Department of Neurosurgery, Brain Tumor Center, Erasmus Medical Center Cancer Institute, Rotterdam, The Netherlands. [email protected].
PMID: 40425813 DOI: 10.1038/s41698-025-00946-1
Abstract

To discover new treatment options for high-grade glioma (HGG), robust in vitro models are essential, but reliably establishing patient-derived cell cultures remains challenging. We established glioma stem-like cell (GSC) cultures from 114 consecutive HGG specimens via traditional surgical resection and/or ultrasonic aspiration, using completely dissociated single cell (single cell-derived, SCD) and partially dissociated 3D-derived (3DD) tissue fragments. Higher success rates in culture establishment were obtained from ultrasonic aspirates and 3DD surgical samples. Combining these approaches yielded a 96% success rate. Copy number profiling showed overall genetic similarities between cultures and parental tissue. Single-cell Sequencing revealed greater transcriptomic heterogeneity in ultrasonic aspiration-derived cultures. Our protocol enabled the screening of 20 anti-cancer agents within a clinically relevant timeframe for 16 out of 18 HGG samples. This refined protocol serves as a robust tool for establishing HGG cell cultures that retain the molecular characteristics of the tumors and support applications in precision medicine.

Products