Gene expression of enhanced green fluorescent protein in immortalized neural progenitor cells strain of rat

    GAO Feng£¬ TIAN Yuª²Ke, YANG Hui, AN Ke£¬ZHANG Chuanª²Han

    Department of Anesthesiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science & Technology, Wuhan 430030, China

    ¡¾Abstract¡¿ AIM: To study the transfection efficiency and the expression of enhanced green fluorescent protein(eGFP) in immortalized neural progenitor cells(INPCs) mediated by recombinant adenoª²associated virus(rAAV). METHODS: The viral vector of rAAV containing eGFP was transfected into INPC. The expression of green fluorescent protein was observed since 24 h after transfection and the transfection efficiency was measured. Antiª²nestin antibodies and simian virus 40 large T antigen gene (SV40Tag) antibodies were used to identify the transfected cells. The specific molecular markers of neurons and astrocytes were detected using immunocytochemistry method to investigate the capability of differentiation of the transfected cells after being induced by 50 mL/L fetal bovine serum. RESULTS: EGFP expression was detected as early as 24 h after transfection. The number of eGFP positive cells reached about 90% after 72 h. After being cultured for 8 weeks, 70%-75% INPCs were still eGFP positive. The transfected cells were confirmed as nestin positive and SV40Tag positive cells with immunocytochemistry and could be differentiated into neurons and astrocytes by the induction of fetal bovine serum. CONCLUSION: The viral vector of rAAV containing eGFP can be highly transfected into INPCs and the transfected cells show a stable and longª²term eGFP expression in vitro. EGFP can be a valuable tracking tool for the study of neural progenitor cell transplantation. ......