A Heterotypic Tridimensional Model to Study the Interaction of Macrophages and Glioblastoma In Vitro
Background: Glioblastoma multiforme (GBM) is the most frequent and aggressive primary brain tumor, and macrophages account for 30–40% of its composition. Most of these macrophages derive from bone marrow monocytes playing a crucial role in tumor progression. Unraveling the mechanisms of macrophages-...
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| Format: | Article |
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MDPI AG
2021-05-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/22/10/5105 |
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| author | María José Gattas Ivana Gisele Estecho María Amparo Lago Huvelle Andrea Emilse Errasti Eugenio Antonio Carrera Silva Marina Simian |
| author_facet | María José Gattas Ivana Gisele Estecho María Amparo Lago Huvelle Andrea Emilse Errasti Eugenio Antonio Carrera Silva Marina Simian |
| author_sort | María José Gattas |
| collection | DOAJ |
| description | Background: Glioblastoma multiforme (GBM) is the most frequent and aggressive primary brain tumor, and macrophages account for 30–40% of its composition. Most of these macrophages derive from bone marrow monocytes playing a crucial role in tumor progression. Unraveling the mechanisms of macrophages-GBM crosstalk in an appropriate model will contribute to the development of specific and more successful therapies. We investigated the interaction of U87MG human GBM cells with primary human CD14<sup>+</sup> monocytes or the THP-1 cell line with the aim of establishing a physiologically relevant heterotypic culture model. Methods: primary monocytes and THP-1 cells were cultured in the presence of U87MG conditioned media or co-cultured together with previously formed GBM spheroids. Monocyte differentiation was determined by flow cytometry. Results: primary monocytes differentiate to M2 macrophages when incubated with U87MG conditioned media in 2-dimensional culture, as determined by the increased percentage of CD14<sup>+</sup>CD206<sup>+</sup> and CD64<sup>+</sup>CD206<sup>+</sup> populations in CD11b<sup>+</sup> cells. Moreover, the mitochondrial protein p32/gC1qR is expressed in monocytes exposed to U87MG conditioned media. When primary CD14<sup>+</sup> monocytes or THP-1 cells are added to previously formed GBM spheroids, both invade and establish within them. However, only primary monocytes differentiate and acquire a clear M2 phenotype characterized by the upregulation of CD206, CD163, and MERTK surface markers on the CD11b<sup>+</sup>CD14<sup>+</sup> population and induce alterations in the sphericity of the cell cultures. Conclusion: our results present a new physiologically relevant model to study GBM/macrophage interactions in a human setting and suggest that both soluble GBM factors, as well as cell-contact dependent signals, are strong inducers of anti-inflammatory macrophages within the tumor niche. |
| first_indexed | 2024-03-10T11:29:21Z |
| format | Article |
| id | doaj.art-084d23ac3b824c91b63e1de8ab6721ec |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T11:29:21Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-084d23ac3b824c91b63e1de8ab6721ec2023-11-21T19:20:11ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-05-012210510510.3390/ijms22105105A Heterotypic Tridimensional Model to Study the Interaction of Macrophages and Glioblastoma In VitroMaría José Gattas0Ivana Gisele Estecho1María Amparo Lago Huvelle2Andrea Emilse Errasti3Eugenio Antonio Carrera Silva4Marina Simian5Instituto de Nanosistemas, Universidad Nacional de San Martín, 25 de Mayo 1021, San Martín, Buenos Aires 1650, ArgentinaInstituto de Farmacología, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155, Piso 9, Buenos Aires 1121, ArgentinaInstituto de Nanosistemas, Universidad Nacional de San Martín, 25 de Mayo 1021, San Martín, Buenos Aires 1650, ArgentinaInstituto de Farmacología, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155, Piso 9, Buenos Aires 1121, ArgentinaInstituto de Medicina Experimental (IMEX), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Academia Nacional de Medicina, Pacheco de Melo 3081, Buenos Aires 1425, ArgentinaInstituto de Nanosistemas, Universidad Nacional de San Martín, 25 de Mayo 1021, San Martín, Buenos Aires 1650, ArgentinaBackground: Glioblastoma multiforme (GBM) is the most frequent and aggressive primary brain tumor, and macrophages account for 30–40% of its composition. Most of these macrophages derive from bone marrow monocytes playing a crucial role in tumor progression. Unraveling the mechanisms of macrophages-GBM crosstalk in an appropriate model will contribute to the development of specific and more successful therapies. We investigated the interaction of U87MG human GBM cells with primary human CD14<sup>+</sup> monocytes or the THP-1 cell line with the aim of establishing a physiologically relevant heterotypic culture model. Methods: primary monocytes and THP-1 cells were cultured in the presence of U87MG conditioned media or co-cultured together with previously formed GBM spheroids. Monocyte differentiation was determined by flow cytometry. Results: primary monocytes differentiate to M2 macrophages when incubated with U87MG conditioned media in 2-dimensional culture, as determined by the increased percentage of CD14<sup>+</sup>CD206<sup>+</sup> and CD64<sup>+</sup>CD206<sup>+</sup> populations in CD11b<sup>+</sup> cells. Moreover, the mitochondrial protein p32/gC1qR is expressed in monocytes exposed to U87MG conditioned media. When primary CD14<sup>+</sup> monocytes or THP-1 cells are added to previously formed GBM spheroids, both invade and establish within them. However, only primary monocytes differentiate and acquire a clear M2 phenotype characterized by the upregulation of CD206, CD163, and MERTK surface markers on the CD11b<sup>+</sup>CD14<sup>+</sup> population and induce alterations in the sphericity of the cell cultures. Conclusion: our results present a new physiologically relevant model to study GBM/macrophage interactions in a human setting and suggest that both soluble GBM factors, as well as cell-contact dependent signals, are strong inducers of anti-inflammatory macrophages within the tumor niche.https://www.mdpi.com/1422-0067/22/10/5105glioblastoma multiformemonocytesmacrophage polarization3D culturestumor-stromal interactionsCD206 |
| spellingShingle | María José Gattas Ivana Gisele Estecho María Amparo Lago Huvelle Andrea Emilse Errasti Eugenio Antonio Carrera Silva Marina Simian A Heterotypic Tridimensional Model to Study the Interaction of Macrophages and Glioblastoma In Vitro International Journal of Molecular Sciences glioblastoma multiforme monocytes macrophage polarization 3D cultures tumor-stromal interactions CD206 |
| title | A Heterotypic Tridimensional Model to Study the Interaction of Macrophages and Glioblastoma In Vitro |
| title_full | A Heterotypic Tridimensional Model to Study the Interaction of Macrophages and Glioblastoma In Vitro |
| title_fullStr | A Heterotypic Tridimensional Model to Study the Interaction of Macrophages and Glioblastoma In Vitro |
| title_full_unstemmed | A Heterotypic Tridimensional Model to Study the Interaction of Macrophages and Glioblastoma In Vitro |
| title_short | A Heterotypic Tridimensional Model to Study the Interaction of Macrophages and Glioblastoma In Vitro |
| title_sort | heterotypic tridimensional model to study the interaction of macrophages and glioblastoma in vitro |
| topic | glioblastoma multiforme monocytes macrophage polarization 3D cultures tumor-stromal interactions CD206 |
| url | https://www.mdpi.com/1422-0067/22/10/5105 |
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