【摘要】 将SV40大T抗原的基因 (LTAg)转染大鼠原代成纤维细胞并筛选到永生化成纤维细胞 (RFLT) ,将此细胞皮下植入裸鼠和大鼠脑内均无致瘤性 ,植入脑内时LTAg基因即停止表达 ,3个月后取出细胞培养时LTAg基因又会重新表达。将RFLT细胞分别转入酪氨酸羟化酶 (TH)、GTP环水解酶 1(GCH)的基因 ,筛选出稳定表达株。混合培养的这两种细胞经HPLC ECD法测出多巴胺 (DA) ,将它们移植入帕金森病 (PD)大鼠模型可明显改善动物的旋转行为 (近 4个月 ) ,在脑切片中观察到TH和增强型绿色荧光蛋白 (EGFP)基因的表达产物。本细胞株的建立为人类PD的基因治疗提供了一种获得新的适用的效应细胞的方法更多还原

【Abstract】 Primary rat fibroblast cells were immortalized by genetic modification of SV40 large T antigen ( LT Ag ) gene and called RFLT. This cell line was non tumorigenic after grafting into nud mouse and rat. The LT Ag gene stopped expression when the cells were transplanted in rat striatum, but it resumed the expression ability after the transplanted cells were recovered from striatum and cultured in vitro. TH gene and GCH gene were transfected into RFLT, respectively, and two types of transfected cells, RFLT TH and RFLT GCH, were obtained. In mixed culture with these two cell lines, DA was detected with HPLC ECD. Implanting mixture of those cells into the striatum of PD rats significantly decreased their rotational asymmetry for up to 12 weeks. The expression of TH gene was proved by TH immunohistochemical staining in the sections of rat brain. The establishment of the genetically modified immortalized cells may play role in the gene therapy of PD.更多还原