兰州大学机构库 >学院待认领
兔骨髓间充质干细胞在神经元增殖与修复中作用的实验研究
Alternative Titleexperiment study on neuron proliferation and repairing effect of bone marrow mesenchymal stem cells on rabbit
李强
Thesis Advisor张建生 ; 丁永忠
2005-05-20
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword骨髓间充质干细胞条件培养基 骨髓间充质干细胞 神经元
Abstract国内外很多研究发现骨髓间充质干细胞(Bone marrow mesenchymal stem cells, BMMSCs)脑内移植能够治疗中枢神经系统疾患,本课题主要通过体外的细胞试验研究,进一步探讨BMMSCs在神经元增殖与修复中的作用。 目的:研究BMMSCs诱导分化为神经样细胞的最佳方案,探讨骨髓间充质干细胞条件培养基(BMMSCs conditioned medium, B-CM)诱导神经干细胞分化为神经细胞的可行性及B-CM对损伤神经元的修复作用。 方法:30只1天~3月龄、体重0.125~1.5kg的青紫蓝兔用于本研究。经过密度梯度离心骨髓分离培养获得BMMSCs,体外扩增传代培养。分别用ATP、GM1、碱性成纤维生长因子,ATP、GM1分别和碱性成纤维生长因子联合诱导BMMSCs。当BMMSCs铺满瓶底90%时,无血清培养24小时,收集细胞培养的上清液为B-CM。体外分离培养兔纹状体神经干细胞,并用条件培养基诱导其分化。用新生兔脊髓神经元建立谷氨酸诱导的神经元损伤模型,观察B-CM损伤神经元的保护作用,用MTT法和LDH法分别检测细胞活性的变化。 结果:BMMSCs特异性表面标记CD44表达阳性,表面标记CD34表达阴性;单纯GM1和ATP不能诱导BMMSCs分化为神经样细胞,碱性成纤维细胞生长因子能单独诱导BMMSCs分化为神经样细胞,神经营养剂联合碱性成纤维细胞生长因子能显著提高诱导分化为神经细胞的比率;从兔纹状体获得呈巢状悬浮生长的细胞团,B-CM能提高其诱导分化率。B-CM能提高损伤神经元的细胞活性。 结论:BMMSCs能促进神经元的增殖与修复。
Other AbstractMany research suggested that central nervous system disorders could be treated by transplantation of bone marrow mesenchymal stem cells(BMMSCs) in vivo. Further research on effects of bone marrow mesenchymal stem cells on neuron proliferation and repairing effect is to be performed in this study. Objectives: To investigate the best method of inducing rabbit BMMSCs to differentiate into neuron-like cells,the possibility of bone marrow mesenchymal stem cells inducing neural stem cells to differentiate into neural cells in conditioned-medium (B-CM)and the effects of B-CM on the damaged neuron repairing. Methods: 30 qingzilan rabbits aged 1 day to 3 months with an average weight of 0.125~1.5kg were used in this study. BMMSCs were harvested from bone marrow and separated using density gradient centrifuge method. BMMSCs cultured and expanded in vitro were induced with gangliosides(GM1)、adenosine triphosphate(ATP)and basic fibroblast growth factor (bFGF ) respectively, and then induced with GM1 /bFGF and ATP/ bFGF. The cultured medium was removed when BMMSCs grew to 90% confluence and then BMMSCs were cultured with serum-free DMEM for 24h and the medium was collected as BMMSCs conditioned-medium(B-CM). Neural stem cells(NSCs) were isolated from the stratium of rabbit and induced by B-CM. The protective effects of B-CM on spinal cord neurons damaged by glutamate were investigated in rabbits. The cells’ viability and repairing degree were assayed by LDH and MMT methods. Results: The antigen CD44 positively expressed but antigen CD34 negatively expressed on BMMSCs. GM1 and ATP couldn’t induce BMMSCs into neural-like cells but bFGF could do that alone. Further more, GM1 /bFGF and ATP/bFGF promote the differentiation rate respectively. The differentiation rate of nest-like clusters of neural stem cells obtained from stratium of rabbits was increased and the viability of damaged neurons was promoted by B-CM. Conclusion: The proliferation and repairing of neurons could be improved by BMMSCs.
URL查看原文
Language中文
Document Type学位论文
Identifierhttp://ir.lzu.edu.cn/handle/262010/222620
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
李强. 兔骨髓间充质干细胞在神经元增殖与修复中作用的实验研究[D]. 兰州. 兰州大学,2005.
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