CT1重组腺病毒转染神经元的培养及鉴定

Culture and identification of neurons transfected with recombinant adenovirus cardiotrophin-1

目的培养、鉴定转染重组腺病毒心肌营养素1(recombinant adenovirus cardiotrophin-1,Adv-CT1)的神经元细胞。方法分离、培养新生Wistar大鼠神经元细胞,将细胞分为Adv-CT1转染组和正常对照组;利用细胞免疫组化技术鉴定CT1基因转染神经元细胞的转染率;利用台盼蓝染色技术检测神经元细胞死亡率。结果 Adv-CT1转染神经元细胞后12、24、36、48及60h转染率分别为43%、75%、90%、95%及97%,相同时间点的死亡率分别为0.12%、0.31%、0.45%、0.48%及0.52%,明显低于正常对照组(0.20%、0.52%、0.83%、1.14%及1.32%)。结论 CT1基因可在体外培养的神经元细胞内持续稳定表达,在转染后48h达到理想转染率,并可降低神经元死亡率,对神经元具有保护作用。

Objective To cultivate and identify the neurons transfected with recombinant adenovirus cardiotrophin-1(Adv-CT1).Methods Hippocampal neurons were isolated and cultured from new born Wistar rats and divided into two groups: Adv-CT1 and control group.The immunocytochemistry technique was used to test the neuron transfection efficiency.The trypanblau staining method was used to test the neuron mortality rate.Results In 12,24,36,48 and 60 hours after transfected with Adv-CT1,transfection rate of neurons was 43%,75%,90%,95% and 97%,respectively,and at the same times,the death rate of neurons was significantly lower in those transfected with Adv-CT1 than that in control group.Conclusions CT1 gene could be expressed steadily in neurons,in vitro,the ideal transfection rate was achieved at 48 hours.Transfected with Adv-CT1 could protect the neurons form damage.