Rat Microglia Cells: Their Culture, Isolation and Phagocytic Activity
| Main Authors: | Fatma, Dewi; Department of Oral Biology, Faculty of Dentistry, Universitas Indonesia, Jakarta 10430, Nakanishi, Hiroshi; Laboratory of Oral Aging Science Faculty of Dental Sciences, Kyushu University Japan |
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| Format: | Article info application/pdf eJournal |
| Bahasa: | eng |
| Terbitan: |
Faculty of Dentistry, University of Indonesia
, 2015
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| Subjects: | |
| Online Access: |
http://www.jdentistry.ui.ac.id/index.php/JDI/article/view/852 http://www.jdentistry.ui.ac.id/index.php/JDI/article/view/852/756 |
Daftar Isi:
- 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 107per 300 cm2 and maintained at 37°C in a 10% Co2/90% 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.