We have recently characterized Neurosphere-derived cells from embryonic day (ED) 17 rat fetus brain. Neural stem cells (NSCs) have been shown to reside in both fetal and adult brain leading to formation of cell clusters termed "neurospheres" and the neurosphere-derived cells can differentiate into neurons, oligodendrocytes and astrocytes. In this study, we reported the establishment of neuronal stem cell line using SV40 large T antigen. The results show that the cells were acquired extended life span and constant increase at early (∼30 cumulative population doubling level (cpdl)), middle (∼60 cpdl), and late (∼100 cpdl) passages. To characterize the established immortal cell, immunofluorescent staining was performed using GFAP antibody for astrocyte, NSE for neuron, and Nestin for neuronal stem cells. These results showed that the established immortal cell line expressed nestin. These results suggest that the established cell line might be a neuronal stem cells, and stem cells might be also major target cells for the carcinogenesis.

Keywords: Immortalization, Stem cells, SV40 Large T-antigen, Nestin, Neurosphere- derived cells