التفاصيل البيبلوغرافية
| العنوان: |
Concomitant Isolation of Primary Astrocytes and Microglia for Protozoa Parasite Infection. |
| المؤلفون: |
Pacheco AOL; Centro de Terapia Celular e Molecular (CTCMol), Universidade Federal de São Paulo (UNIFESP)., Amaral MP; Centro de Terapia Celular e Molecular (CTCMol), Universidade Federal de São Paulo (UNIFESP)., de Farias IS; Centro de Terapia Celular e Molecular (CTCMol), Universidade Federal de São Paulo (UNIFESP)., Bottino LZMF; Centro de Terapia Celular e Molecular (CTCMol), Universidade Federal de São Paulo (UNIFESP)., Bortoluci KR; Departamento de Ciências Biológicas, Centro de Terapia Celular e Molecular (CTCMol), Universidade Federal de São Paulo (UNIFESP); kbortoluci@gmail.com. |
| المصدر: |
Journal of visualized experiments : JoVE [J Vis Exp] 2020 Mar 18 (157). Date of Electronic Publication: 2020 Mar 18. |
| نوع المنشور: |
Journal Article; Research Support, Non-U.S. Gov't; Video-Audio Media |
| اللغة: |
English |
| بيانات الدورية: |
Publisher: MYJoVE Corporation Country of Publication: United States NLM ID: 101313252 Publication Model: Electronic Cited Medium: Internet ISSN: 1940-087X (Electronic) Linking ISSN: 1940087X NLM ISO Abbreviation: J Vis Exp Subsets: MEDLINE |
| أسماء مطبوعة: |
Original Publication: [Boston, Mass. : MYJoVE Corporation, 2006]- |
| مواضيع طبية MeSH: |
Astrocytes/*cytology , Microglia/*cytology , Parasitic Diseases/*pathology, Animals ; Animals, Newborn ; Astrocytes/parasitology ; Cell Culture Techniques ; Cerebral Cortex/cytology ; Cerebral Cortex/parasitology ; Mice ; Microglia/parasitology ; Parasitic Diseases/parasitology ; Toxoplasma/physiology ; Trypanosoma cruzi/physiology |
| مستخلص: |
Astrocytes and microglia are the most abundant glial cells. They are responsible for physiological support and homeostasis maintenance in the central nervous system (CNS). The increasing evidences of their involvement in the control of infectious diseases justify the emerging interest in the improvement of methodologies to isolate primary astrocytes and microglia in order to evaluate their responses to infections that affect the CNS. Considering the impact of Trypanosoma cruzi (T. cruzi) and Toxoplasma gondii (T. gondii) infection in the CNS, here we provide a method to extract, maintain, dissociate and infect murine astrocytes and microglia cells with protozoa parasites. Extracted cells from newborn cortices are maintained in vitro for 14 days with periodic differential media replacement. Astrocytes and microglia are obtained from the same extraction protocol by mechanical dissociation. After phenotyping by flow cytometry, cells are infected with protozoa parasites. The infection rate is determined by fluorescence microscopy at different time points, thus enabling the evaluation of differential ability of glial cells to control protozoan invasion and replication. These techniques represent simple, cheap and efficient methods to study the responses of astrocytes and microglia to infections, opening the field for further neuroimmunology analysis. |
| تواريخ الأحداث: |
Date Created: 20200407 Date Completed: 20200916 Latest Revision: 20200916 |
| رمز التحديث: |
20221213 |
| DOI: |
10.3791/60680 |
| PMID: |
32250348 |
| قاعدة البيانات: |
MEDLINE |
