Observation of differentiation and ultrastructure of neural progenitor cells from embryonic rat hippocampus in vitro
AIM: To explore the methods of isolation and culture of neural progenitor cells (NPCs) from hippocampus of embryonic rats and to observe the characteristics of induced differentiation and ultrastructure of NPCs. METHODS: Primary NPCs were isolated from Wistar rat (E15 - 16 d) hippocampus tissues and the cells were suspended and cultured in serum free medium (containing bFGF and EGF) for long-term survival in vitro, and were induced to differentiate in medium containing fetal calf serum. Immunocytochemical staining was applied to identify the NPCs and their potential of differentiation and electron microscopes were employed to observe the ultrastructure of the differentiated cells. RESULTS: The cells isolated from hippocampus of embryonic rats could be continuously passage-cultured in vitro and form clone. They were nestin positive. The cells after differentiation expressed the specific antigens β Ⅲ tubulin (tuj-1) and glial fibrillary acidic protein (GFAP) in neurons and astrocytes, respectively. Nervous processes formed network and various kinds of developed organelles structure were observed in differentiated cells under electron microscopes. Junction-like structures between the cells and intact synapse in some mature differentiated cells were observed. CONCLUSION: These results indicate that the NPCs isolated and cultured using the present methods are capable of proliferation and multi-potential differentiation.
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万方数据
维普
