Scaffold-Based (Matrigel™) 3D Culture Technique of Glioblastoma Recovers a Patient-like Immunosuppressive Phenotype
Conventional 2D cultures are commonly used in cancer research though they come with limitations such as the lack of microenvironment or reduced cell heterogeneity. In this study, we investigated in what respect a scaffold-based (Matrigel™) 3D culture technique can ameliorate the limitations of 2D cu...
| Main Authors: | , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-07-01
|
| Series: | Cells |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2073-4409/12/14/1856 |
| _version_ | 1827733518389608448 |
|---|---|
| author | Frank K. Braun Tanja Rothhammer-Hampl Julia Lorenz Sandra Pohl Ayse-Nur Menevse Arabel Vollmann-Zwerenz Elisabeth Bumes Maren Büttner Saida Zoubaa Martin Proescholdt Nils O. Schmidt Peter Hau Philipp Beckhove Beate Winner Markus J. Riemenschneider |
| author_facet | Frank K. Braun Tanja Rothhammer-Hampl Julia Lorenz Sandra Pohl Ayse-Nur Menevse Arabel Vollmann-Zwerenz Elisabeth Bumes Maren Büttner Saida Zoubaa Martin Proescholdt Nils O. Schmidt Peter Hau Philipp Beckhove Beate Winner Markus J. Riemenschneider |
| author_sort | Frank K. Braun |
| collection | DOAJ |
| description | Conventional 2D cultures are commonly used in cancer research though they come with limitations such as the lack of microenvironment or reduced cell heterogeneity. In this study, we investigated in what respect a scaffold-based (Matrigel™) 3D culture technique can ameliorate the limitations of 2D cultures. NGS-based bulk and single-cell sequencing of matched pairs of 2D and 3D models showed an altered transcription of key immune regulatory genes in around 36% of 3D models, indicating the reoccurrence of an immune suppressive phenotype. Changes included the presentation of different HLA surface molecules as well as cellular stressors. We also investigated the 3D tumor organoids in a co-culture setting with tumor-infiltrating lymphocytes (TILs). Of note, lymphocyte-mediated cell killing appeared less effective in clearing 3D models than their 2D counterparts. IFN-γ release, as well as live cell staining and proliferation analysis, pointed toward an elevated resistance of 3D models. In conclusion, we found that the scaffold-based (Matrigel™) 3D culture technique affects the transcriptional profile in a subset of GBM models. Thus, these models allow for depicting clinically relevant aspects of tumor-immune interaction, with the potential to explore immunotherapeutic approaches in an easily accessible in vitro system. |
| first_indexed | 2024-03-11T01:12:26Z |
| format | Article |
| id | doaj.art-34a1a8f1e65a4f44afca380bf5cfda63 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-11T01:12:26Z |
| publishDate | 2023-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-34a1a8f1e65a4f44afca380bf5cfda632023-11-18T18:46:09ZengMDPI AGCells2073-44092023-07-011214185610.3390/cells12141856Scaffold-Based (Matrigel™) 3D Culture Technique of Glioblastoma Recovers a Patient-like Immunosuppressive PhenotypeFrank K. Braun0Tanja Rothhammer-Hampl1Julia Lorenz2Sandra Pohl3Ayse-Nur Menevse4Arabel Vollmann-Zwerenz5Elisabeth Bumes6Maren Büttner7Saida Zoubaa8Martin Proescholdt9Nils O. Schmidt10Peter Hau11Philipp Beckhove12Beate Winner13Markus J. Riemenschneider14Department of Neuropathology, Regensburg University Hospital, 93053 Regensburg, GermanyDepartment of Neuropathology, Regensburg University Hospital, 93053 Regensburg, GermanyDepartment of Neuropathology, Regensburg University Hospital, 93053 Regensburg, GermanyDepartment of Neuropathology, Regensburg University Hospital, 93053 Regensburg, GermanyDivision of Interventional Immunology, Leibniz Institute for Immunotherapy, 93053 Regensburg, GermanyDepartment of Neurology and Wilhelm Sander-NeuroOncology Unit, Regensburg University Hospital, 93053 Regensburg, GermanyDepartment of Neurology and Wilhelm Sander-NeuroOncology Unit, Regensburg University Hospital, 93053 Regensburg, GermanyInstitute of Computational Biology, Helmholtz Center Munich, 85764 Munich, GermanyDepartment of Neuropathology, Regensburg University Hospital, 93053 Regensburg, GermanyDepartment of Neurosurgery, Regensburg University Hospital, 93053 Regensburg, GermanyDepartment of Neurosurgery, Regensburg University Hospital, 93053 Regensburg, GermanyDepartment of Neurology and Wilhelm Sander-NeuroOncology Unit, Regensburg University Hospital, 93053 Regensburg, GermanyDivision of Interventional Immunology, Leibniz Institute for Immunotherapy, 93053 Regensburg, GermanyDepartment of Stem Cell Biology, Friedrich-Alexander-Universität (FAU) Erlangen-Nürnberg, 91054 Erlangen, GermanyDepartment of Neuropathology, Regensburg University Hospital, 93053 Regensburg, GermanyConventional 2D cultures are commonly used in cancer research though they come with limitations such as the lack of microenvironment or reduced cell heterogeneity. In this study, we investigated in what respect a scaffold-based (Matrigel™) 3D culture technique can ameliorate the limitations of 2D cultures. NGS-based bulk and single-cell sequencing of matched pairs of 2D and 3D models showed an altered transcription of key immune regulatory genes in around 36% of 3D models, indicating the reoccurrence of an immune suppressive phenotype. Changes included the presentation of different HLA surface molecules as well as cellular stressors. We also investigated the 3D tumor organoids in a co-culture setting with tumor-infiltrating lymphocytes (TILs). Of note, lymphocyte-mediated cell killing appeared less effective in clearing 3D models than their 2D counterparts. IFN-γ release, as well as live cell staining and proliferation analysis, pointed toward an elevated resistance of 3D models. In conclusion, we found that the scaffold-based (Matrigel™) 3D culture technique affects the transcriptional profile in a subset of GBM models. Thus, these models allow for depicting clinically relevant aspects of tumor-immune interaction, with the potential to explore immunotherapeutic approaches in an easily accessible in vitro system.https://www.mdpi.com/2073-4409/12/14/1856brain cancertumor organoidsGBMnext-generation sequencingsingle-cell RNA sequencing |
| spellingShingle | Frank K. Braun Tanja Rothhammer-Hampl Julia Lorenz Sandra Pohl Ayse-Nur Menevse Arabel Vollmann-Zwerenz Elisabeth Bumes Maren Büttner Saida Zoubaa Martin Proescholdt Nils O. Schmidt Peter Hau Philipp Beckhove Beate Winner Markus J. Riemenschneider Scaffold-Based (Matrigel™) 3D Culture Technique of Glioblastoma Recovers a Patient-like Immunosuppressive Phenotype Cells brain cancer tumor organoids GBM next-generation sequencing single-cell RNA sequencing |
| title | Scaffold-Based (Matrigel™) 3D Culture Technique of Glioblastoma Recovers a Patient-like Immunosuppressive Phenotype |
| title_full | Scaffold-Based (Matrigel™) 3D Culture Technique of Glioblastoma Recovers a Patient-like Immunosuppressive Phenotype |
| title_fullStr | Scaffold-Based (Matrigel™) 3D Culture Technique of Glioblastoma Recovers a Patient-like Immunosuppressive Phenotype |
| title_full_unstemmed | Scaffold-Based (Matrigel™) 3D Culture Technique of Glioblastoma Recovers a Patient-like Immunosuppressive Phenotype |
| title_short | Scaffold-Based (Matrigel™) 3D Culture Technique of Glioblastoma Recovers a Patient-like Immunosuppressive Phenotype |
| title_sort | scaffold based matrigel™ 3d culture technique of glioblastoma recovers a patient like immunosuppressive phenotype |
| topic | brain cancer tumor organoids GBM next-generation sequencing single-cell RNA sequencing |
| url | https://www.mdpi.com/2073-4409/12/14/1856 |
| work_keys_str_mv | AT frankkbraun scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT tanjarothhammerhampl scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT julialorenz scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT sandrapohl scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT aysenurmenevse scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT arabelvollmannzwerenz scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT elisabethbumes scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT marenbuttner scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT saidazoubaa scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT martinproescholdt scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT nilsoschmidt scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT peterhau scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT philippbeckhove scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT beatewinner scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype AT markusjriemenschneider scaffoldbasedmatrigel3dculturetechniqueofglioblastomarecoversapatientlikeimmunosuppressivephenotype |