Refined culture conditions with increased physiological relevance uncover oncogene-dependent metabolic signatures in Ewing sarcoma spheroids
Ewing sarcoma (EwS) cell line culture largely relies on standard techniques, which do not recapitulate physiological conditions. Here, we report on a feasible and cost-efficient EwS cell culture technique with increased physiological relevance employing an advanced medium composition, reduced fetal calf serum, and spheroidal growth. Improved reflection of the transcriptional activity related to proliferation, hypoxia, and differentiation in EwS patient tumors was detected in EwS cells grown in this refined in vitro condition. Moreover, transcriptional signatures associated with the oncogenic activity of the EwS-specific FET::ETS fusion transcription factors in the refined culture condition were shifted from proliferative toward metabolic gene signatures. The herein-presented EwS cell culture technique with increased physiological relevance provides a broadly applicable approach for enhanced in vitro modeling relevant to advancing EwS research and the validity of experimental results.
Authors
- Florencia Cidre‐Aranaz (ORCID: https://orcid.org/0000-0002-0246-7179)
- Martha J. Carreño-Gonzalez (ORCID: https://orcid.org/0000-0003-2718-9002)
- Florian H. Geyer
- Shunya Ohmura (ORCID: https://orcid.org/0000-0002-0930-5172)
- A. Katharina Ceranski (ORCID: https://orcid.org/0000-0001-6754-4190)
- Tobias Faehling (ORCID: https://orcid.org/0000-0002-5500-4987)
- Anna C. Ehlers
- Kimberley M. Hanssen (ORCID: https://orcid.org/0000-0002-2919-3104)
- Nadine Gmelin
- Philipp Poeller
- Almut Schulze (ORCID: https://orcid.org/0000-0002-8199-6422)
- Endrit Vinca (ORCID: https://orcid.org/0009-0005-1920-290X)
- Zuzanna Kolodynska
- Thomas G.P. Grünewald
Institutions
- German Cancer Research Center (DE)
- Heidelberg University (DE)
- University Hospital Heidelberg (DE)
- National Center for Tumor Diseases (DE)
- Hopp Children's Cancer Center Heidelberg (DE)
- DKFZ-ZMBH Alliance (DE)
- Deutsches Konsortium für Translationale Krebsforschung (DE)
Publication Details
- Journal
- Cell Reports Methods
- Published
- 2025-02-01
- DOI
- https://doi.org/10.1016/j.crmeth.2025.100966
- Citations
- 6
- Primary Topic
- Sarcoma Diagnosis and Treatment
- Type
- article
- Field-Weighted Citation Impact
- 9.11
Funders
- Deutsches Krebsforschungszentrum
- Matthias Lackas-Stiftung
- European Commission
- Deutsche Forschungsgemeinschaft
- Bundesministerium für Bildung und Forschung
- Deutsche Krebshilfe
- Stichting Kinderen Kankervrij
- Deutschen Konsortium für Translationale Krebsforschung
- Dr. Rolf M. Schwiete Stiftung
- European Research Council