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Nucleofection of Rodent Neuroblasts to Study Neuroblast Migration In vitro
Published on: November 12, 2013
Scanning electron microscopic study of human neuroblastoma cells affected with Naegleria fowleri Thai strains
Supathra Tiewcharoen1, Jundee Rabablert, Pruksawan Chetanachan
1Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, 2 Prannok Rd., Bangkok, 10700, Thailand.
Abstract:
In order to understand the pathogenesis of Naegleria fowleri in primary amoebic meningoencephalitis, the human neuroblastoma (SK-N-MC) and African green monkey kidney (Vero) cells were studied in vitro. Amoeba suspension in cell-culture medium was added to the confluent monolayer of SK-N-MC and Vero cells. The cytopathic activity of N. fowleri trophozoites in co-culture system was elucidated by scanning electron microscope at 3, 6, 9, 12, and 24 h. Two strains of N. fowleri displayed well-organized vigorous pseudopods in Nelson's medium at 37 degrees C. In co-culture, the target monolayer cells were damaged by two mechanisms, phagocytosis by vigorous pseudopods and engulfment by sucker-like apparatus. N. fowleri trophozoites produced amoebostomes only in co-culture with SK-N-MC cells. In contrast, we could not find such apparatus in the co-culture with Vero cells. The complete destruction time (100%) at 1:1 amoeba/cells ratio of SK-N-MC cells (1 day) was shorter than the Vero cells (12 days). In conclusion, SK-N-MC cells were confirmed to be a target model for studying neuropathogenesis of primary amoebic meningoencephalitis.
