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: | Book |
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Faculty of Dentistry, Universitas Indonesia,
2015-10-01T00:00:00Z.
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| Summary: | <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> |
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| Item Description: | 1693-9697 2355-4800 10.14693/jdi.v12i1.852 |