摘要
目的从人脐血中分离单个核细胞(MNCs)、体外培养扩增间充质干细胞(MSCs)并研究其生物学特性和定向诱导能力。方法无菌条件下采集脐血,肝素抗凝,分离单个核细胞。用DMEM/F12培养基进行纯化和扩增培养,分别向神经元样细胞诱导,获取MSCs,显微镜下观察它的形态、尼氏体染色;取扩增2、5、7代的MSCs,免疫细胞化学法检测nestin表达和神经元样细胞特异性标记。结果诱导扩增后的MSCs尼氏体染色阳性,第2、5、7代MSCs的nestin的阳性表达率分别为(51.2±3.2)%、(34.6±2.7)%、(11.3±3.3)%;神经元特异性烯醇化酶(NSE)在原代细胞无表达,而在2、5、7代MSC的阳性表达率分别为(11.4±2.3)%、(21.78±3.1)%、(40.7±3.4)%。结论人脐血MSCs具有神经干/祖细胞的特性,在一定条件下能向神经元样细胞分化。
Objective The mononuclear cells （MNCs） were cultivated and expanded into mesenchymal stem cells （MSCs） from human umbilical cord blood, and the purpose of this study was to explore the biological characteristics and induced differentiation ability in vitro. Methods Human umbilical cord blood samples were obtained and the mononuclear cells were isolated from it, then inoculated the MNCs into 25-mm culture flasks containing DMEM/F12 medium. The morphology was observed under microscope. Nissl body staining was used, The passage 2, 4, 7 of the expanded MSCs were induced to differentiate to neuron-like cells. The expressions of nestin and neuron-specific enolase （NSE） on the treated cells were detected by immunocytochemical method. Results Nissl body staining was positive ; Nestin expression was found in （51.2 ± 3.2） % of the second, （34. 6± 2. 7 ） % of the fifth, （ 11.3 ± 3.3 ） % of the seventh passage of MSCs; NSE expression was found in （ 11.4 ± 2. 3 ） % of the second, （21.78 ± 3. 1 ） % of the fifth, （40. 7 ± 3.4） % of the seventh passage of MSCs. Conclusion Cord blood MSCs possess some features of neural stem cells, and have the capacity to differentiate into neuron-like cells under proper conditions.
出处
《基础医学与临床》
CSCD
北大核心
2007年第3期298-301,共4页
Basic and Clinical Medicine
基金
河南省医学科技创新人才工程项目(2005018)
关键词
脐血
间充质干细胞
神经元样细胞
诱导分化
cord blood
mesenchymal stem cells
neuron-like cell
induction differentiation
