{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T11:18:09Z","timestamp":1783423089303,"version":"3.54.6"},"reference-count":22,"publisher":"IMR Press","issue":"6","license":[{"start":{"date-parts":[[2026,6,22]],"date-time":"2026-06-22T00:00:00Z","timestamp":1782086400000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Front. Biosci. (Landmark Ed)"],"abstract":"<jats:p>Background:Most data related to high-grade glioma and glioblastoma (GBM) 3D models refer to ultra-low\/suspension cultivation systems, for several reasons: easy to initiate, high penetrability of treatments, ease of harvesting for downstream analysis. However, the major limitation of these models is the absence of the extracellular matrix, which enables the specific, local invasiveness of these aggressive brain tumors.Aim:This study provides a comparison between ultra-low and several extracellular matrix (ECM) and hydrogel culture conditions for various grades of patient-derived glioma organoids, in terms of cell growth, invasiveness, and release of cytokines\/chemokines, as a model for downstream analysis of soluble factors.Materials and Methods:Patient-derived glioma organoids were initiated in three different conditions: ultra-low attachment(UL), 3D Matrigel, and dextran-based hydrogels. Human endothelial cells were used for tube assay co-cultures and human normal monocyte cell line SC to assess monocyte invasion and interaction with tumor organoids.Results:UL conditions supported organoid formation with better growth rate and easy passaging, and reliable detection of secreted cytokines and chemokines, closely mirroring qualitative secretory profiles observed in tumor explants. In contrast, ECM-based systems enabled invasive growth patterns and co-culture assays, but with a restricted immune cell infiltration and with a reduced detectable level of soluble factors. While qualitative cytokine signatures were preserved across culture platforms, substantial quantitative differences were observed, highlighting the influence of matrix composition on measurable outputs. Preliminary data showed that organoid-forming capacity indicated no clear association with Isocitrate Dehydrogenase 1 (IDH1) mutational status in the tested samples.Conclusion:Our findings indicate that UL and ECM-based models provide complementary information and recommend a dual-platform cell culture initiation strategy to maximize robustness and translational relevance in patient-derived glioma research.<\/jats:p>","DOI":"10.31083\/fbl51339","type":"journal-article","created":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T10:50:51Z","timestamp":1783421451000},"source":"Crossref","is-referenced-by-count":0,"title":["Comparative Analysis of Ultra-Low Versus Hydrogel Growing Conditions for Patient-Derived Glioma Organoids"],"prefix":"10.31083","volume":"31","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5071-9756","authenticated-orcid":true,"given":"Maria","family":"Dud\u0103u","sequence":"first","affiliation":[{"name":"Biochemistry-Proteomics Department, Victor Babes National Institute of Pathology, 050096 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mugurel Petrinel","family":"Radoi","sequence":"additional","affiliation":[{"name":"Neurovascular Surgery Department, Carol Davila University of Medicine and Pharmacy, 050047 Bucharest, Romania"},{"name":"Department of Vascular Neurosurgery, National Institute of Neurology and Neurovascular Diseases, 077160 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ionela Daniela","family":"Popescu","sequence":"additional","affiliation":[{"name":"Biochemistry-Proteomics Department, Victor Babes National Institute of Pathology, 050096 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicoleta","family":"Constantin","sequence":"additional","affiliation":[{"name":"Biochemistry-Proteomics Department, Victor Babes National Institute of Pathology, 050096 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valeriu Bogdan","family":"Cismasiu","sequence":"additional","affiliation":[{"name":"Cell Biology, Neurosciences and Experimental Miology Department, Victor Babes National Institute of Pathology, 050096 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"S\u0103voaia","sequence":"additional","affiliation":[{"name":"Cellular and Molecular Biology and Histology Department, Carol Davila University of Medicine and Pharmacy, 050047 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8122-1096","authenticated-orcid":true,"given":"Ana-Maria","family":"Enciu","sequence":"additional","affiliation":[{"name":"Biochemistry-Proteomics Department, Victor Babes National Institute of Pathology, 050096 Bucharest, Romania"},{"name":"Cellular and Molecular Biology and Histology Department, Carol Davila University of Medicine and Pharmacy, 050047 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2122-0235","authenticated-orcid":true,"given":"Cristiana","family":"Tanase","sequence":"additional","affiliation":[{"name":"Biochemistry-Proteomics Department, Victor Babes National Institute of Pathology, 050096 Bucharest, Romania"},{"name":"Faculty of Medicine, Titu Maiorescu University, 031593 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5507-1020","authenticated-orcid":true,"given":"Elena","family":"Codrici","sequence":"additional","affiliation":[{"name":"Biochemistry-Proteomics Department, Victor Babes National Institute of Pathology, 050096 Bucharest, Romania"},{"name":"Cellular and Molecular Biology and Histology Department, Carol Davila University of Medicine and Pharmacy, 050047 Bucharest, Romania"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"16242","published-online":{"date-parts":[[2026,6,22]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","unstructured":"Greenwald AC, Darnell NG, Hoefflin R, Simkin D, Mount CW, Gonzalez Castro LN, et al. Integrative spatial analysis reveals a multi-layered organization of glioblastoma. Cell. 2024; 187: 2485\u20132501.e26.","DOI":"10.1016\/j.cell.2024.03.029"},{"key":"ref2","doi-asserted-by":"publisher","unstructured":"Jacob F, Salinas RD, Zhang DY, Nguyen PTT, Schnoll JG, Wong SZH, et al. A Patient-Derived Glioblastoma Organoid Model and Biobank Recapitulates Inter- and Intra-tumoral Heterogeneity. Cell. 2020; 180: 188\u2013204.e22.","DOI":"10.1016\/j.cell.2019.11.036"},{"key":"ref3","doi-asserted-by":"publisher","unstructured":"Park J, Kim D, Park HC, Park M, Zhang S, Na O, et al. Tumor microenvironment-preserving gliosarcoma organoids as an in vitro preclinical platform: a comparative analysis with glioblastoma models. Journal of Translational Medicine. 2025; 23: 915.","DOI":"10.1186\/s12967-025-06952-y"},{"key":"ref4","doi-asserted-by":"publisher","unstructured":"Chen JWE, Pedron S, Harley BAC. The Combined Influence of Hydrogel Stiffness and Matrix-Bound Hyaluronic Acid Content on Glioblastoma Invasion. Macromolecular Bioscience. 2017; 17:  https:\/\/doi.org\/10.1002\/mabi.201700018","DOI":"10.1002\/mabi.201700018"},{"key":"ref5","doi-asserted-by":"publisher","unstructured":"Ruiz-Garcia H, Alvarado-Estrada K, Schiapparelli P, Quinones-Hinojosa A, Trifiletti DM. Engineering Three-Dimensional Tumor Models to Study Glioma Cancer Stem Cells and Tumor Microenvironment. Frontiers in Cellular Neuroscience. 2020; 14: 558381.","DOI":"10.3389\/fncel.2020.558381"},{"key":"ref6","doi-asserted-by":"publisher","unstructured":"Del Roc\u00edo Aguilera-Marquez J, Manzanares-Guzm\u00e1n A, Garc\u00eda-Uriostegui L, Canales-Aguirre AA, Camacho-Villegas TA, Lugo-Fabres PH. Alginate-Gelatin Hydrogel Scaffold Model for Hypoxia Induction in Glioblastoma Embedded Spheroids. Gels (Basel, Switzerland). 2025; 11: 263.","DOI":"10.3390\/gels11040263"},{"key":"ref7","doi-asserted-by":"publisher","unstructured":"Liang L, Cui R, Zhong S, Wang Z, He Z, Duan H, et al. Analysis of the potential role of photocurable hydrogel in patient-derived glioblastoma organoid culture through RNA sequencing. Biomaterials Science. 2022; 10:4902\u20134914.","DOI":"10.1039\/d2bm00589a"},{"key":"ref8","doi-asserted-by":"publisher","unstructured":"Birocchi F, Cusimano M, Rossari F, Beretta S, Rancoita PMV, Ranghetti A, et al. Targeted inducible delivery of immunoactivating cytokines reprograms glioblastoma microenvironment and inhibits growth in mouse models. Science Translational Medicine. 2022; 14: eabl4106.","DOI":"10.1126\/scitranslmed.abl4106"},{"key":"ref9","doi-asserted-by":"publisher","unstructured":"Rossari F, Birocchi F, Naldini L, Coltella N. Gene-based delivery of immune-activating cytokines for cancer treatment. 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Annals of Oncology. 2022; 33: S1411.","DOI":"10.1016\/j.annonc.2022.09.091"},{"key":"ref13","doi-asserted-by":"publisher","unstructured":"McCoy MG, Nyanyo D, Hung CK, Goerger JP, R. Zipfel W, Williams RM, et al. Endothelial cells promote 3D invasion of GBM by IL-8-dependent induction of cancer stem cell properties. Scientific Reports. 2019; 9: 9069.","DOI":"10.1038\/s41598-019-45535-y"},{"key":"ref14","doi-asserted-by":"publisher","unstructured":"Shah N, Hallur PM, Ganesh RA, Sonpatki P, Naik D, Chandrachari KP, et al. Gelatin methacrylate hydrogels culture model for glioblastoma cells enriches for mesenchymal-like state and models interactions with immune cells. Scientific Reports. 2021; 11: 17727.","DOI":"10.1038\/s41598-021-97059-z"},{"key":"ref15","doi-asserted-by":"publisher","unstructured":"Park J, Koh I, Cha J, Oh Y, Shim JK, Kim H, et al. Comparison of Glioblastoma Cell Culture Platforms Based on Transcriptional Similarity with Paired Tissue. Pharmaceuticals (Basel, Switzerland). 2024; 17: 529.","DOI":"10.3390\/ph17040529"},{"key":"ref16","doi-asserted-by":"publisher","unstructured":"Majc B, Habi\u010d A, Malavolta M, Vittori M, Por\u010dnik A, Bo\u0161njak R, et al. Patient-derived tumor organoids mimic treatment-induced DNA damage response in glioblastoma. iscience. 2024; 27: 110604.","DOI":"10.1016\/j.isci.2024.110604"},{"key":"ref17","doi-asserted-by":"publisher","unstructured":"Lindemann C, Marschall V, Weigert A, Klingebiel T, Fulda S. Smac Mimetic-Induced Upregulation of CCL2\/MCP-1 Triggers Migration and Invasion of Glioblastoma Cells and Influences the Tumor Microenvironment in a Paracrine Manner. Neoplasia (New York, N.Y.). 2015; 17: 481\u2013489.","DOI":"10.1016\/j.neo.2015.05.002"},{"key":"ref18","doi-asserted-by":"publisher","unstructured":"Migliorini E, Thakar D, K\u00fchnle J, Sadir R, Dyer DP, Li Y, et al. Cytokines and growth factors cross-link heparan sulfate. Open Biology. 2015; 5: 150046.","DOI":"10.1098\/rsob.150046"},{"key":"ref19","doi-asserted-by":"publisher","unstructured":"Fujita M, Ishihara M, Ono K, Hattori H, Kurita A, Shimizu M, et al. Adsorption of inflammatory cytokines using a heparin-coated extracorporeal circuit. Artificial Organs. 2002; 26: 1020\u20131025.","DOI":"10.1046\/j.1525-1594.2002.07017.x"},{"key":"ref20","doi-asserted-by":"publisher","unstructured":"Na MK, Oh Y, Lee D, Park J, Yoon SJ, Yoo J, et al. Comparison of the biological characteristics of glioblastoma tumorspheres obtained from fresh and cryopreserved glioblastoma tissues. Journal of Neuro-Oncology. 2025; 174: 191\u2013206.","DOI":"10.1007\/s11060-025-05052-7"},{"key":"ref21","doi-asserted-by":"publisher","unstructured":"Zhang Y, Shao Y, Li Y, Li X, Zhang X, E Q, et al. The generation of glioma organoids and the comparison of two culture methods. Cancer Medicine. 2024; 13: e7081.","DOI":"10.1002\/cam4.7081"},{"key":"ref22","doi-asserted-by":"publisher","unstructured":"Singh SK, Abbas S, Saxena AK, Tiwari S, Sharma LK, Tiwari M. Critical role of three-dimensional tumorsphere size on experimental outcome. 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