CAJ | 학술논문

目的检测神经干细胞经谷氨酸脱羧酶(GAD65)基因转染并诱导分化后γ-氨基丁酸(cABA)的表达,为GAD65基因修饰神经干细胞移植治疗癫痫提供理论依据.方法采用新一代高效脂质体作为转染试剂的基因转导方法转染神经干细胞,通过免疫细胞荧光方法鉴定神经干细胞分化为GABA阳性神经元情况.结果采用脂质体转染方法能成功地将CAD60基因转染至神经干细胞中,转染的神经干细胞在分化第7天及第14天GABA阳性神经元分别占(31.9±4.2)%、(26.5±2.3)%,未转染的神经干细胞分化的GABA阳性神经元则分別占(28.4±1.9)%和(20.3±3.5)%,转染者与未转染者相比,分化的GABA阳性神经元百分比有显著性差异(P<0.05).结论采用脂质体基因转染技术,可以将GAD65基因转染至神经干细胞中,并促进神经干细胞分化为GABA神经元.
목적 검측신경간세포경곡안산탈최매(GAD65)기인전염병유도분화후γ-안기정산(cABA)적표체,위GAD65기인수식신경간세포이식치료전간제공이론의거.방법 채용신일대고효지질체작위전염시제적기인전도방법전염신경간세포,통과면역세포형광방법감정신경간세포분화위GABA양성신경원정황.결과 채용지질체전염방법능성공지장CAD60기인전염지신경간세포중,전염적신경간세포재분화제7천급제14천GABA양성신경원분별점(31.9±4.2)%、(26.5±2.3)%,미전염적신경간세포분화적GABA양성신경원칙분별점(28.4±1.9)%화(20.3±3.5)%,전염자여미전염자상비,분화적GABA양성신경원백분비유현저성차이(P<0.05).결론 채용지질체기인전염기술,가이장GAD65기인전염지신경간세포중,병촉진신경간세포분화위GABA신경원.
Objectives To sbldy the expression of Gamma-aminobutyric acid (GABA) by neural stem cells (NSCs) transfected with dutamic acid decarboxylase (GAD65) gene in order to provide the basis for treatment of epilepsy by implanting NSCs transfected with GAD65 gene.Methods NSCs were transfected by PcDNA3-GAD65 liposomes which is a new generation of highly efficient transfection reagent.The GABA-positive neurons differentited by transfected NSCs with PcDNA3-GAD65 and non-transfected NSCs was identified by the method of immune cells fluorescent. Results NSCs were successfully transfected by PcDNA3-GAD65. The quantities of GABA-positive neutons were(31.9±4.2)% and (26.5±2.3)% respectively in the first 7 days and 14 days of dlfferentiation in the transfected group, and (28.4±1.9)% and (20.3±3.5)% respectively in the non-transfected group. There was a significant differenct in the quantity of GABA-positive neurons between both the transfected and non-transfected groups (P<O.05). Conclusion GAD65 gene can be successfully transfected to the NSCs by the liposomes and neurons differentiated from the GAD65 gene-modified NSCs can secrete a numbers of GABA.