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中国人民解放军总医院
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中国人民解放军总医院老年心血管病研究所
中国科技出版传媒股份有限公司
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中华老年多器官疾病杂志编辑委员会
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E-mail: zhlndqg@mode301.cn
创刊人 王士雯
总编辑 范利
副总编辑 陈韵岱
执行主编 叶大训
编辑部主任 王雪萍
ISSN 1671-5403
CN 11-4786
创刊时间 2002年
出版周期 月刊
邮发代号 82-408
友情链接
人尿源干细胞的分离培养及体外成神经分化研究
In Vitro Culture and Neural Differentiation of Human Urine-derived Stem Cells
投稿时间:2018-11-01  修订日期:2018-12-24
DOI:
中文关键词:  人尿源干细胞;神经干细胞;神经胶质细胞;分化
英文关键词:Human urine-drived stem cells, neuron cell, neuroglia cell, differentiation
基金项目:国家自然科学基金面上项目(81570808);上海市卫生系统优秀人才(2017BR045);国家重点研发计划(2017YFC0906903)
作者单位E-mail
姜之歆 第六人民医院内分泌代谢科室 jiangzhixin1992@163.com 
王从容 第六人民医院内分泌代谢科室 crwang@sjtu.edu.cn 
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中文摘要:
      目的 从正常成人尿液中分离提取间充质干细胞(MSC)来源的人尿源干细胞(hUSC),并研究比较其在不同诱导液中的体外成神经分化能力。方法 提取正常成人来源hUSC,经体外培养、扩增,流式鉴定后,用4种不同配方的成神经分化液分别诱导hUSC向神经细胞分化,在诱导至第14天时,光镜下观察细胞形态改变, 并用RT-PCR方法进行分化能力鉴定。结果 从正常成人提取的hUSC成功表达MSC表面标志物(CD27+, CD44+, CD73+, CD90+, CD31-, CD34-, CD45-, HLA-DR-)。hUSC在B、C、D组成神经分化液诱导培养14天后,细胞胞体回缩饱满、折光度变强,呈现神经元样突起。RT-PCR 结果诱导14天后,A组各标志物均未检测出;B组、C组、D组神经干细胞(NSC)特异性标志物Nestin的相对表达量均显著增高(Ps < 0.01),其中C组诱导液增高最为突出(95.63±3.79,P < 0.01);B组、C组、D组诱导分化后胶质细胞特异性前体标志物S100的表达量也显著增高(Ps < 0.01),其中B组诱导液效果较好(6.15±0.18, P < 0.01);但成熟神经元细胞特异性标志物β3-tublin与NF-200基因表达量并未增加。结论 hUSC具备MSC特性;B、C、D组分化液均有助于hUSC向神经类细胞方向分化,提示在优化诱导条件下,hUSC能向NSC及胶质细胞亚群分化。
英文摘要:
      Objective Human urinary stem cells (hUSCs) with mesenchymal stem cells (MSCs) characteristics were isolated from urine of healthy individuals, and its neurogenic differentiation capabilities in different neurogenic induction mediums were investigated. Methods HUSCs from normal individuals were extracted, cultured, and identified. Four different neurogenic differentiation methods were used. After induction for 14 days, cell morphology was observed under the microscope, and differentiation capability was identified by RT-PCR. Results HUSCs extracted from healthy adults successfully expressed MSCs surface markers (CD27+, CD44+, CD73+, CD90+, CD31-, CD34-, CD45-, HLA-DR-). After 14 days induction in B, C, D neural differentiation medium, hUSCs showed a retraction of the cell body and high refracting power, as well as processing neuron-like protuberance. However, RT-PCR showed relative markers were not detectable in medium A after 14 days induction. The expression of Nestin, a specific marker of neural stem cells (NSCs), increased significantly in medium B、C、D (Ps < 0.01), especially in medium C(95.63±3.79); The expression of S100, a specific marker of percursor cells of oligodendrocytes and astrocytes also increased in medium B、C、D (Ps < 0.01), among which the effect of medium B was better(6.15±0.18); The expression of neuron cell-specific markers β3-tublin and NF-200 were not increased in all medium. Conclusion HUSCs possessed MSCs characteristics. The B, C, D neural differentiation medium contributed to the differentiation of hUSCs into neuron cells. The results suggest that hUSC can differentiate into NSC and glial subpopulations under optimized induction conditions.
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