A Pro-Inflammatory Stimulus versus Extensive Passaging of DITNC1 Astrocyte Cultures as Models to Study Astrogliosis
Abstract
:1. Introduction
2. Results
2.1. DITNC1 Astrocytes with Extensive Passaging but Not Low Passage Respond to Neuronal Thy-1 in the Absence of Inflammatory Stimuli
2.2. Cells with a Low Number of Passages Behave as Non-Reactive Astrocytes and Switch to Reactive Astrocytes When Treated with TNF
2.3. Reactive Phenotype Traits of DITNC1 Cells with Extensive Passaging or Treated with Pro-Inflammatory Stimuli Are Not Due to Senescence
2.4. DITNC1 Astrocytes with Multiple Passages Inhibit Neurite Outgrowth and Promote Neuronal Death
3. Discussion
4. Materials and Methods
4.1. Cell Culture
4.2. Reagents and Antibodies
4.3. Immunoblot Analysis
4.4. Cell Cytometry
4.5. Migration Assay in a Boyden Chamber
4.6. Indirect Immunofluorescence Assay for Visualization of Focal Adhesions
4.7. Proliferation Assays
4.8. Beta-Galactosidase Senescence Assay
4.9. Cell Cycle Assay
4.10. Outgrowth of Neuronal Processes on Fixed Astrocyte Monolayers
4.11. Neuronal Cell Death Assay
4.12. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
References
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Pérez, L.A.; Palacios, E.; González, M.F.; Leyton-Rivera, I.; Martínez-Meza, S.; Pérez-Núñez, R.; Jeldes, E.; Avalos, A.M.; Díaz, J.; Leyton, L. A Pro-Inflammatory Stimulus versus Extensive Passaging of DITNC1 Astrocyte Cultures as Models to Study Astrogliosis. Int. J. Mol. Sci. 2024, 25, 9454. https://doi.org/10.3390/ijms25179454
Pérez LA, Palacios E, González MF, Leyton-Rivera I, Martínez-Meza S, Pérez-Núñez R, Jeldes E, Avalos AM, Díaz J, Leyton L. A Pro-Inflammatory Stimulus versus Extensive Passaging of DITNC1 Astrocyte Cultures as Models to Study Astrogliosis. International Journal of Molecular Sciences. 2024; 25(17):9454. https://doi.org/10.3390/ijms25179454
Chicago/Turabian StylePérez, Leonardo A., Esteban Palacios, María Fernanda González, Ignacio Leyton-Rivera, Samuel Martínez-Meza, Ramón Pérez-Núñez, Emanuel Jeldes, Ana María Avalos, Jorge Díaz, and Lisette Leyton. 2024. "A Pro-Inflammatory Stimulus versus Extensive Passaging of DITNC1 Astrocyte Cultures as Models to Study Astrogliosis" International Journal of Molecular Sciences 25, no. 17: 9454. https://doi.org/10.3390/ijms25179454