Protocol for in vitro analysis of pro-inflammatory and metabolic functions of cultured primary murine astrocytes
Summary: Robust protocols are required to investigate in vitro the molecular mechanisms that control astrocyte metabolism and pro-inflammatory activities. In the present protocol, we describe step by step the isolation and culture of primary murine astrocytes from neonatal brains, followed by their...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-03-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721007395 |
| _version_ | 1818321129467543552 |
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| author | Cristina Gutiérrez-Vázquez Francisco J. Quintana |
| author_facet | Cristina Gutiérrez-Vázquez Francisco J. Quintana |
| author_sort | Cristina Gutiérrez-Vázquez |
| collection | DOAJ |
| description | Summary: Robust protocols are required to investigate in vitro the molecular mechanisms that control astrocyte metabolism and pro-inflammatory activities. In the present protocol, we describe step by step the isolation and culture of primary murine astrocytes from neonatal brains, followed by their genetic manipulation with siRNA. We further describe cytokine activation of the cultured astrocytes for the analysis of their pro-inflammatory responses, and the oxygen consumption analysis to assess their metabolic function.For complete details on the use and execution of this protocol, please refer to Chao et al. (2019), Clark et al. (2021), and Rothhammer et al. (2018). |
| first_indexed | 2024-12-13T10:36:00Z |
| format | Article |
| id | doaj.art-5bc357b4aa124a6b9586b45009355ca2 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-13T10:36:00Z |
| publishDate | 2022-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-5bc357b4aa124a6b9586b45009355ca22022-12-21T23:50:43ZengElsevierSTAR Protocols2666-16672022-03-0131101033Protocol for in vitro analysis of pro-inflammatory and metabolic functions of cultured primary murine astrocytesCristina Gutiérrez-Vázquez0Francisco J. Quintana1Ann Romney Center for Neurologic Diseases, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USA; Corresponding authorAnn Romney Center for Neurologic Diseases, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Corresponding authorSummary: Robust protocols are required to investigate in vitro the molecular mechanisms that control astrocyte metabolism and pro-inflammatory activities. In the present protocol, we describe step by step the isolation and culture of primary murine astrocytes from neonatal brains, followed by their genetic manipulation with siRNA. We further describe cytokine activation of the cultured astrocytes for the analysis of their pro-inflammatory responses, and the oxygen consumption analysis to assess their metabolic function.For complete details on the use and execution of this protocol, please refer to Chao et al. (2019), Clark et al. (2021), and Rothhammer et al. (2018).http://www.sciencedirect.com/science/article/pii/S2666166721007395Cell cultureCell isolationImmunologyMetabolismNeuroscience |
| spellingShingle | Cristina Gutiérrez-Vázquez Francisco J. Quintana Protocol for in vitro analysis of pro-inflammatory and metabolic functions of cultured primary murine astrocytes STAR Protocols Cell culture Cell isolation Immunology Metabolism Neuroscience |
| title | Protocol for in vitro analysis of pro-inflammatory and metabolic functions of cultured primary murine astrocytes |
| title_full | Protocol for in vitro analysis of pro-inflammatory and metabolic functions of cultured primary murine astrocytes |
| title_fullStr | Protocol for in vitro analysis of pro-inflammatory and metabolic functions of cultured primary murine astrocytes |
| title_full_unstemmed | Protocol for in vitro analysis of pro-inflammatory and metabolic functions of cultured primary murine astrocytes |
| title_short | Protocol for in vitro analysis of pro-inflammatory and metabolic functions of cultured primary murine astrocytes |
| title_sort | protocol for in vitro analysis of pro inflammatory and metabolic functions of cultured primary murine astrocytes |
| topic | Cell culture Cell isolation Immunology Metabolism Neuroscience |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721007395 |
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