摘要
目的 制备胶质细胞源性神经生长因子 ( GDNF)基因修饰的骨髓基质干细胞 ( MSCs) ,观察其对多巴胺能神经元的作用 ,探索治疗帕金森氏病的新途径。方法 应用逆转录聚合酶链反应 ( RT- PCR)方法从新生小鼠大脑皮层细胞克隆出 GDNF c DNA片断 ,以 p EGFP- C1为载体导入 MSCs,制备稳定表达 GDNF基因的 MSCs工程细胞 ,采用联合培养的技术通过倒置显微镜和免疫组织化学的方法观察 MSCs和 GDNF基因修饰的 MSCs工程细胞与多巴胺能神经元之间的相互作用。结果 MSCs和 GDNF基因修饰的 MSCs工程细胞均能促进多巴胺能神经元的存活和生长 ,MSCs工程细胞作用更强。结论 成功构建了 GDNF基因修饰的 MSCs工程细胞 ,该细胞对多巴胺能神经元有明显营养保护作用 。
Objective To construct a kind of engineered cell secreting GDNF,observe its protective effect on primary dopaminergic neurons and study its possible effects on gene therapy of Parkinson's disease.Methods Mouse GDNF cDNA was amplified from newly-born mouse cortex cell of cerebrumby RT-PCR,and ligated into the pEGFP-C1 vector.Then transferring the recombined plasmid into MSCs and screening engineered MSCs which can secrete GDNF protein stably.Dopaminergic neuron cells were co-cultured with MSCs and MSCs which modified by GDNF gene respectively,the effects between these cells were observed using phase-contrast microscope and immunohistochemistry staining.Results Both MSCs and GDNF-modified MSCs showed great neurotrophic and protective effections on dopaminergic neurons,but the effections of GDNF-modified MSCs were stronger than MSCs.Conlusions The GDNF-modified MSCs have been constructed successfully.These cells obviously promoted the survival and growth of dopaminergic neurons.
出处
《中风与神经疾病杂志》
CAS
CSCD
北大核心
2004年第4期311-313,i001,共4页
Journal of Apoplexy and Nervous Diseases
