成年大鼠骨髓间充质干细胞可塑性的体外研究
发布时间：2019-05-15 12:04
【摘要】： 第一部分骨髓间充质干细胞的多向分化潜能 第一章体外诱导成年大鼠骨髓间充质干细胞分化为5羟色胺敏感性神经元 目的:体外诱导成年大鼠骨髓间充质干细胞分化为具有神经元表型和部分功能的细胞。 方法:将取自6-8周成年SD大鼠股、胫骨的细胞贴壁传代培养至6-10代,采用有限稀释法对细胞作单克隆纯化。在全反式视黄酸预诱导24小时后,HBSS洗去含血清培养液,换用改良的Woodbury长时程神经元诱导液培养细胞,3、6、9、12、24、48、72、96小时后显微镜下观察细胞形态变化;细胞免疫染色检测神经元标志物神经元特异性烯醇化酶、神经丝M+H、5-羟色胺的表达;并用胞内钙代谢测定法测定诱导前后细胞对5-HT的功能反应。 结果:改良神经元诱导液处理3小时后,细胞开始表现神经元的形态特征,细胞折光性增强,形成收缩的双极或多极胞体和细长突起。细胞可以维持神经元样存活72小时以上。诱导5小时后,对免疫染色
[Abstract]:The first part is the multidirectional differentiation potential of bone marrow mesenchymal stem cells. Chapter 1 inducing adult rat bone marrow mesenchymal stem cells to differentiate into serotonin sensitive neurons in vitro Aim: to induce adult rat bone marrow mesenchymal stem cells to differentiate into cells with neuronal phenotype and partial function in vitro. Methods: the cells taken from the strands of adult SD rats for 6 weeks and 8 weeks were cultured to 6 脳 10 generations, and the cells were purified by finite dilution method. After 24 hours of all-trans Retinic acid pretreatment, HBSS washed away the serum culture medium and replaced it with the modified Woodbury long-term neuron induction solution. The cells were cultured in 3, 6, 9, 12, 24, 48, 72, 96 hours, and the morphological changes of the cells were observed under microscope. The expression of neuron-specific enolase and 5-hydroxytryptamine in nerve filament was detected by cellular immunostaining, and the functional response of cells to 5-HT before and after induction was measured by intracellular calcium metabolism assay. Results: after 3 hours of modified neuron induction solution treatment, the cells began to show the morphological characteristics of neurons, the refractive index of the cells was enhanced, and the contractile bipolar or multipolar cell bodies and slender processes were formed. Cells can maintain neuron-like survival for more than 72 hours. After 5 hours of induction, immunostaining
【学位授予单位】：重庆医科大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R329
本文编号：2477489
[Abstract]:The first part is the multidirectional differentiation potential of bone marrow mesenchymal stem cells. Chapter 1 inducing adult rat bone marrow mesenchymal stem cells to differentiate into serotonin sensitive neurons in vitro Aim: to induce adult rat bone marrow mesenchymal stem cells to differentiate into cells with neuronal phenotype and partial function in vitro. Methods: the cells taken from the strands of adult SD rats for 6 weeks and 8 weeks were cultured to 6 脳 10 generations, and the cells were purified by finite dilution method. After 24 hours of all-trans Retinic acid pretreatment, HBSS washed away the serum culture medium and replaced it with the modified Woodbury long-term neuron induction solution. The cells were cultured in 3, 6, 9, 12, 24, 48, 72, 96 hours, and the morphological changes of the cells were observed under microscope. The expression of neuron-specific enolase and 5-hydroxytryptamine in nerve filament was detected by cellular immunostaining, and the functional response of cells to 5-HT before and after induction was measured by intracellular calcium metabolism assay. Results: after 3 hours of modified neuron induction solution treatment, the cells began to show the morphological characteristics of neurons, the refractive index of the cells was enhanced, and the contractile bipolar or multipolar cell bodies and slender processes were formed. Cells can maintain neuron-like survival for more than 72 hours. After 5 hours of induction, immunostaining
【学位授予单位】：重庆医科大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R329
【参考文献】
相关期刊论文 前3条
1 项鹏,夏文杰,张丽蓉,陈振光,张秀明,李艳,李树浓;成人骨髓间质干细胞定向诱导为神经元样细胞的研究[J];中国病理生理杂志;2001年05期
2 许念桂,肖波,葛亮,杨国帅,曹贵方;钙离子在颞叶癫沲大鼠海马神经元内的动态变化[J];中华神经科杂志;2005年10期
3 卓本慧,江和碧,瞿平,李廷玉;骨髓间充质干细胞向神经细胞定向分化的体外研究[J];中国修复重建外科杂志;2005年05期
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