人脐带间充质干细胞的体外扩增及向神经元样细胞定向诱导分化

发布时间：2018-01-22 01:35

  本文关键词： 脐带 间充质干细胞 神经细胞 分化 诱导 丹参　出处：《汕头大学》2006年硕士论文　论文类型：学位论文

【摘要】：目的：研究人脐带间充质干细胞(MSCs)的体外分离、纯化、扩增及丹参诱导MSCs向神经元样细胞分化，，为神经细胞移植、脐带组织库的建立及中药丹参作为神经诱导剂提供理论依据。方法：无菌条件下收集剖宫产新生儿脐带，组织块培养法获取MSCs，进行培养。丹参诱导人脐带来源的MSCs向神经细胞分化；用免疫细胞化学方法检测神经细胞相关蛋白标记的表达；用光镜、扫描电子显微镜监测丹参诱导后细胞形态改变；透射电子显微镜观察丹参诱导后细胞超微结构改变；用共聚焦显微镜分析丹参诱导前后细胞内DNA、RNA含量改变及神经干细胞的标记巢蛋白(nestin)，神经元的蛋白标记β-Ⅲ类神经微管(β-TubulinⅢ)表达含量变化。用RT-PCR检测细胞神经相关基因pleiotrophin和神经干细胞的标记巢蛋白(nestin)的表达。RT-PCR检测证实，经丹参诱导后的MSCs表达神经干细胞相关基因Nestin，诱导前和诱导后的MSCs均表达神经细胞基因Pleiotrophin，诱导后的表达明显增强。结果：从人脐带华尔通胶分离、培养的贴壁细胞，体外生长形态类似于成纤维细胞，可以维持在未分化状态稳定增殖，体外增殖超过10代，细胞冻存一个月后
[Abstract]:Objective: to study the isolation, purification, amplification and differentiation of MSCs into neuron-like cells from human umbilical cord mesenchymal stem cells (MSCs) in vitro. The establishment of umbilical cord tissue bank and traditional Chinese medicine Salvia miltiorrhiza as a nerve inducer to provide theoretical basis. Methods: the umbilical cord of newborn born by cesarean section was collected under aseptic condition and MSCs was obtained by tissue mass culture. Salvia miltiorrhiza induced the differentiation of MSCs from human umbilical cord into neural cells. The expression of neuron-associated protein was detected by immunocytochemistry. The morphological changes of cells induced by Salvia miltiorrhiza were monitored by light microscope and scanning electron microscope. The ultrastructure of cells induced by Salvia miltiorrhiza was observed by transmission electron microscope. The changes of DNA RNA content and the labeled nestin of neural stem cells were analyzed by confocal microscope before and after salvia miltiorrhiza induction. Protein labeling of Neuron 尾-鈪

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