摘要
目的 构建GDNF基因修饰的骨髓基质干细胞 ,并观察其对多巴胺能神经元的营养支持作用。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GDNFcDNA片段 ,以pEGFP C1为载体导入骨髓基质干细胞 (MSCs) ,制备稳定表达GDNF/EGFP融合基因的MSCs工程细胞 ,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的相互作用。 结果 MSCs和GDNF基因修饰的MSCs工程细胞共培养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 and study the supportive effects of the engineered cells on dopaminergic neuron. Methods Mouse GDNF cDNA was amplified from newly-born mouse cerebrum by RT-PCR,and ligated into the pEGFP-C1 vector. Then the recombined plasmid was transfected into bone marrow stromal cells(MSCs), and engineered MSCs were screened. Embryonic mesencephalic neurons were co-cultured with MSCs that had and had not been modified by GDNF gene, respectively ,and followd by immunohistochemistry staining for tyrosinase hydroxylase. Results Both GDNF-modified and unmodified MSCs showed great neurotrophic and protective effections on dopaminergic neurons with more efficiency of GDNF-modified MSCs than unmodified cells. Conclusion The GDNF-modified MSCs obviously promote the survival and growth of cultured dopaminergic neurons.
