间充质干细胞的条件培养上清对PC12细胞帕金森病模型增殖的影响  

Effect of conditioned supernatant of mesenchymal stem cell on the proliferation of PC12 cell model of Parkinson disease

作　　者：刘旻[1] 刘金[2] 刘新珊 李鹏飞[2] 苏忠玉[2] 李彤[1] 段海峰[2] 

机构地区：[1]新乡医学院第一附属医院,河南卫辉453100 [2]军事医学科学院放射与辐射研究所,北京100850

出　　处：《新乡医学院学报》2015年第12期1064-1067,1072,共5页Journal of Xinxiang Medical University

摘　　要：目的探讨间充质干细胞(MSC)的条件培养上清对PC12细胞及其帕金森病(PD)模型增殖的影响。方法组织块贴壁法从脐带组织中分离出长梭形细胞,用流式细胞术鉴定其免疫表型,用碱性磷酸酶染色法鉴定其分化能力;用刀豆素A(Con A)刺激MSC后收集其条件培养上清,观察MSC条件培养上清与正常MSC上清对PC12及其PD模型细胞增殖能力的影响,采用酶联免疫吸附测定法检测2种上清内粒细胞集落刺激因子(G-CSF)水平。结果经实验分离的MSC高表达干细胞表面标志CD105、CD73、CD90,而不表达造血细胞表面标志CD19、CD34和CD45,符合MSC表型特征。MSC经Con A处理后,表型未发生变化。MSC具有良好的成骨、成脂分化能力,经Con A处理的MSC,其成骨、成脂分化能力未发生明显变化。与正常MSC上清比较,经Con A处理后的MSC条件上清可显著提高PC12细胞及其PD模型细胞的增殖能力(P<0.05)。经过Con A诱导的MSC条件培养上清中G-CSF水平为(32.13±0.31)ng·L-1,未经处理的MSC上清中G-CSF水平为(11.13±0.28)ng·L-1,经过Con A诱导的MSC条件培养上清中G-CSF水平显著高于未经处理的MSC上清(P<0.05)。结论 Con A可促进PC12细胞及其PD模型细胞增殖,G-CSF可能参与了促进PC12细胞及其PD模型细胞增殖的过程。Objective To study the effect of conditioned supernatant of mesenchymal stem cell（ MSC） on the proliferation of PC12 cell and its cell model of Parkinson disease（ PD）. Methods The long fusiform cells were isolated from umbilical tissue by tissue adherent method,the immunophenotype of the cells was identified by flow cytometry,and the differentiation ability was identified by alkaline phosphatase staining method. The MSCs were stimulated by canavalin A（ Con A）,and their conditioned supernatants was collected. The effect of the MSC conditioned supernatant and normal supernatant on reproductive activity of PC12 cells and its cell model of PD. The level of granulocyte colony stimulating factor（ G-CSF） in the two kinds of supernatant was detected by enzyme-linked immunosorbent assay. Results The surface markers CD105,CD73 and CD90 of MSC were high expression; but there was no expression in the hematopoietic cell surface markers CD19,CD34 and CD45; the result was consistent with the MSC phenotype. The phenotype of MSC did not change after treatment with Con A.MSC had good osteogenic and adipogenic differentiation ability,the osteogenic and adipogenic differentiation ability did not significantly change after treatment with Con A. Compared with normal MSC supernatant,the conditioned supernatant of MSC treated with Con A could significantly improve the proliferation ability of PC12 cells and PD cells（ P〈0. 05）. The level of G-CSF in the conditioned supernatant treated with Con A and common supernatant of MSC was（ 32. 13 ± 0. 31） and（ 11. 13 ± 0. 28） ng·L- 1respectively,the level of G-CSF in the conditioned supernatant of MSC treated with Con A was significantly higher than that in the common supernatant of MSC（ P〈0. 05）. Conclusions Con A can promote the proliferation of PC12 cell and its cell model of PD. G-CSF may be involved in the process of promoting the proliferation of PC12 cell and its cell model of PD.

关 键 词：间充质干细胞 PC12细胞 帕金森病 增殖 粒细胞集落刺激因子 

分 类 号：R742.5[医药卫生—神经病学与精神病学]