AccScience Publishing / IJB / Online First / DOI: 10.36922/IJB025520537
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RESEARCH ARTICLE

3D bioprinting of the glioblastoma microenvironment for preclinical assessment of CDK4/6 inhibition

Philipp Kaps1 ,  Emily Zunke1 ,  Justus Ramtke1 ,  Christian Polley2 ,  Leonora Calopresti2 ,  Marcus Frank3,4 ,  Karoline Schulz4 ,  Piotr Grabarczyk5 ,  Sascha Troschke-Meurer6 ,  Charlotte Wagner1 ,  Annabell Wolff1 ,  Daniel Dubinski7 ,  Florian Gessler7 ,  Thomas M. Freiman7 ,  Christian Junghanss1 ,  Hermann Seitz2,3 ,  Claudia Maletzki1*
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1 Department of Internal Medicine–Clinic and Polyclinic for Hematology, Hemostaseology, Oncology, Stem Cell Therapy and Palliative Medicine, Rostock University Medical Center, University of Rostock, Rostock, Germany
2 Faculty of Mechanical Engineering and Marine Technology, University of Rostock, Rostock, Germany
3 Department Life, Light & Matter, University of Rostock, Rostock, Germany
4 Medical Biology and Electron Microscopy Centre, Rostock University Medical Center, University of Rostock, Rostock, Germany
5 Department of Internal Medicine, Clinic III–Hematology, Oncology, University Medicine Greifswald, Greifswald, Germany
6 Department of Pediatric Oncology and Hematology, University Medicine Greifswald, Greifswald, Germany
7 Department of Neurosurgery, Rostock University Medical Center, University of Rostock, Rostock, Germany
IJB, 025520537 https://doi.org/10.36922/IJB025520537
Received: 27 November 2025 | Accepted: 6 January 2026 | Published online: 13 January 2026
© 2026 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
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Abstract

Glioblastoma (GBM) is an aggressive, World Health Organization grade 4 brain tumor with a poor prognosis, largely due to its complex, treatment-resistant microenvironment. To better model this environment for preclinical testing, we developed a three-dimensional (3D) biomimetic bioprinting platform using patient-derived GBM cells. Two hydrogels, alginate/gelatin (AlgGel; 1.5%/7.5%) and gelatin methacryloyl (10%), were evaluated for biocompatibility. GBM cells (GBM06, GBM14, and GBM15), transduced with iRFP-680 for viability tracking, were embedded in the hydrogels and printed. Tumor growth and viability were monitored for 28 days using fluorescence microscopy, complemented by electron microscopy (EM) for structural analysis. Drug response testing included temozolomide (TMZ; 10 μM) and the cyclin-dependent kinases 4/6 inhibitor abemaciclib (1 μM). Cell viability and extracellular vesicle (EV) release were quantified. Efficacy was further assessed in a co-culture with astrocytes. The AlgGel hydrogel supported superior long-term viability and growth. EM analysis of AlgGel scaffolds revealed preserved cellular architecture and adherence to the bioprinted extracellular matrix. Drug response assays confirmed findings previously observed in 2D and 3D cultures. Two cycles of abemaciclib reduced GBM cell viability in AlgGel scaffolds, accompanied by a significant decrease in EV secretion. TMZ, in contrast, did not significantly affect cell viability. The reduction in viability remained pronounced in co-culture with astrocytes, without compromising astrocyte viability. In this study, we present a  3D biomimetic bioprinting model that successfully mimics key aspects of the GBM microenvironment. This model demonstrates strong potential as a preclinical drug screening tool, enabling improved mechanistic insight into cell–matrix interactions that govern nutrient/metabolite diffusion and therapeutic responses.

Graphical abstract
Keywords
Abemaciclib
Alginate–gelatin hydrogel
CDK4/6 inhibition
Cell–matrix interaction
Glioblastoma
Patient-derived tumor cells
Three-dimensional culture
Tumor microenvironment
Funding
The project was funded by the German Federal Institute for Risk Assessment Grant Agreement Number 60- 0102-01.P640 to C.M. and C.P. and by Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – SFB 1270/1,2–299150580 to H.S.
Conflict of interest
The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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International Journal of Bioprinting, Electronic ISSN: 2424-8002 Print ISSN: 2424-7723, Published by AccScience Publishing
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