大鼠神经干细胞体外培养条件下GDNF及NGF的分泌特点  

GDNF and NGF Secretions of Cultured Neural Stem Cells

作　　者：靳晓亮[1] 李京红[2] 关方霞[3] 杜英[4] 樊锐太[1] 

机构地区：[1]郑州大学第一附属医院神经外科,郑州450052 [2]郑州大学第一附属医院神经内科,郑州450052 [3]郑州大学生物工程系,郑州450001 [4]郑州大学基础医学院微生物与免疫学教研室,郑州450052

出　　处：《神经损伤与功能重建》2009年第2期108-109,113,共3页Neural Injury and Functional Reconstruction

摘　　要：目的:探讨体外培养条件下大鼠神经干细胞(NSCs)分泌胶质细胞源性神经营养因子(GDNF)和神经生长因子(NGF)的特点。方法:取SD胎鼠NSCs悬浮培养,光镜观察细胞形态,使用Nestin、微管相关蛋白2(MAP-2)、胶质纤维酸性蛋白(GFAP)、半乳糖神经鞘氨醇酶(GalC)免疫荧光法鉴定细胞。培养第3代后使用ELISA法测量其GDNF及NGF分泌量,连续测量6代。结果:光镜下第3代细胞仍保持较多NSCs特征,至第6代,大部分干细胞分化为星形胶质细胞,少部分分化为神经元或少突胶质细胞。第3代细胞GDNF的分泌量较低,但随着培养时间的延长,其分泌量逐渐增高;而第3代细胞NGF的分泌量较高,但其分泌有一个高峰期(第6代),随后逐渐下降。结论:在体外培养条件下,NSCs干细胞阶段分泌GDNF的能力较低,而分泌NGF的能力较高;进入分化阶段后,分泌GDNF的能力提高,而分泌NGF的能力有所减弱。利用干细胞移植治疗中枢神经系统损伤时应注意时程的选择。Objective：To investigate the status of GDNF and NGF secretions of cultured neural stem cells in vitro. Methods：Neural stem cells （NSCs） of S-D rats were prepared and identified by immunocytoehemistry. The secretions of GDNF and NT 3 were measured by ELISA. Results： Majority of the cells in the third passage were determined as neural stem cells. In the sixth passage, most stem cells differentiated into astroeytes as indicated by the expression of GFAP while few expressed specific markers for neurons or oligodentrocytes. ELISA results have demonstrated that the total content of GDNF in the third passage cells was initially low and gradually increased in the cultured process. However, the total content of NGF was relatively high, and reached a peak in the sixth passage and then decreased. Conclusion：The secretions of GDNF and NGF of the cultured neural stem cells show distinct dynamic patterns in the process of culture. More attention should be paid to the passage choice during NSCs transplantation therapy.

关 键 词：神经干细胞 胶质细胞源性神经营养因子 神经生长因子 

分 类 号：R741[医药卫生—神经病学与精神病学]