Rat Microglia Cells: Their Culture, Isolation and Phagocytic Activity
<p>Microglia were isolated from mixed primary cell cultures of the cerebral cortex from 3day old male Wistar rats. The mechanically dissociated cells were plated in a flask at a density of 10<sup>7</sup>per 300 cm<sup>2</sup> and maintained at 37°C in a 10% Co<sub>...
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| Format: | Article |
| Language: | English |
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Faculty of Dentistry, Universitas Indonesia
2015-10-01
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| Series: | Journal of Dentistry Indonesia |
| Subjects: | |
| Online Access: | http://jdentistry.ui.ac.id/index.php/JDI/article/view/852 |
| _version_ | 1811338478288371712 |
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| author | Dewi Fatma Hiroshi Nakanishi |
| author_facet | Dewi Fatma Hiroshi Nakanishi |
| author_sort | Dewi Fatma |
| collection | DOAJ |
| description | <p>Microglia were isolated from mixed primary cell cultures of the cerebral cortex from 3day old male Wistar rats. The mechanically dissociated cells were plated in a flask at a density of 10<sup>7</sup>per 300 cm<sup>2</sup> and maintained at 37°C in a 10% Co<sub>2</sub>/90% air atmosphere. After 10-14 days in culture, floating and attached cells on the mixed primary cultured cell layer were isolated by gentle shaking of the flask for 5 min. The resulting cell suspension was transferred to plastic dishes and allowed to adhere at 37°C . To investigate the morphological change of microglia, the cells after 2 days of culture were incubated with biotinylated GSA-1-B4 (10µg/ml) at 4°C for overnight. To detect the phagocytic activity, isolated microglia were incubated with opsonized zymosan (20mgl/ml) for Ih at 37°C and with Giemsa's staining solution for 30 min at room temperature. The results were about 90% of attached cells had amoeboid and rod-shaped cell bodies with no or a few thick processes. Most of these cells became amoeboid-like cells and showed a number of vacuoles in the cytosol when cultured in the presence of IFN-ɣ+LPS. Both control and IFN-ɣ + LPS – treated cells exhibited the intense phagocytic activity against zymosan particles.</p> |
| first_indexed | 2024-04-13T18:11:26Z |
| format | Article |
| id | doaj.art-98de0d02d69e4410b1b2ddcbe8de26e4 |
| institution | Directory Open Access Journal |
| issn | 1693-9697 2355-4800 |
| language | English |
| last_indexed | 2024-04-13T18:11:26Z |
| publishDate | 2015-10-01 |
| publisher | Faculty of Dentistry, Universitas Indonesia |
| record_format | Article |
| series | Journal of Dentistry Indonesia |
| spelling | doaj.art-98de0d02d69e4410b1b2ddcbe8de26e42022-12-22T02:35:53ZengFaculty of Dentistry, Universitas IndonesiaJournal of Dentistry Indonesia1693-96972355-48002015-10-011211410.14693/jdi.v12i1.852746Rat Microglia Cells: Their Culture, Isolation and Phagocytic ActivityDewi Fatma0Hiroshi Nakanishi1Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta 10430Laboratory of Oral Aging Science Faculty of Dental Sciences, Kyushu University Japan<p>Microglia were isolated from mixed primary cell cultures of the cerebral cortex from 3day old male Wistar rats. The mechanically dissociated cells were plated in a flask at a density of 10<sup>7</sup>per 300 cm<sup>2</sup> and maintained at 37°C in a 10% Co<sub>2</sub>/90% air atmosphere. After 10-14 days in culture, floating and attached cells on the mixed primary cultured cell layer were isolated by gentle shaking of the flask for 5 min. The resulting cell suspension was transferred to plastic dishes and allowed to adhere at 37°C . To investigate the morphological change of microglia, the cells after 2 days of culture were incubated with biotinylated GSA-1-B4 (10µg/ml) at 4°C for overnight. To detect the phagocytic activity, isolated microglia were incubated with opsonized zymosan (20mgl/ml) for Ih at 37°C and with Giemsa's staining solution for 30 min at room temperature. The results were about 90% of attached cells had amoeboid and rod-shaped cell bodies with no or a few thick processes. Most of these cells became amoeboid-like cells and showed a number of vacuoles in the cytosol when cultured in the presence of IFN-ɣ+LPS. Both control and IFN-ɣ + LPS – treated cells exhibited the intense phagocytic activity against zymosan particles.</p>http://jdentistry.ui.ac.id/index.php/JDI/article/view/852Microglia cellsPrimary cell culturesPhagocytic activity |
| spellingShingle | Dewi Fatma Hiroshi Nakanishi Rat Microglia Cells: Their Culture, Isolation and Phagocytic Activity Journal of Dentistry Indonesia Microglia cells Primary cell cultures Phagocytic activity |
| title | Rat Microglia Cells: Their Culture, Isolation and Phagocytic Activity |
| title_full | Rat Microglia Cells: Their Culture, Isolation and Phagocytic Activity |
| title_fullStr | Rat Microglia Cells: Their Culture, Isolation and Phagocytic Activity |
| title_full_unstemmed | Rat Microglia Cells: Their Culture, Isolation and Phagocytic Activity |
| title_short | Rat Microglia Cells: Their Culture, Isolation and Phagocytic Activity |
| title_sort | rat microglia cells their culture isolation and phagocytic activity |
| topic | Microglia cells Primary cell cultures Phagocytic activity |
| url | http://jdentistry.ui.ac.id/index.php/JDI/article/view/852 |
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