基础医学与临床 ›› 2007, Vol. 27 ›› Issue (3): 298-301.

• 研究论文 • 上一篇    下一篇

人脐血间充质干细胞的分离鉴定及其临床意义

徐虹 阮丽荣 杨波 杜英 胡祥 胡平生 王云龙   

  1. 河南省肿瘤医院肿瘤研究所 郑州大学基础医学院微生物与免疫学教研室 郑州大学基础医学院微生物与免疫学教研室 瑞典Biovitron公司 河南省生物工程研究所
  • 收稿日期:2006-03-17 修回日期:2006-08-04 出版日期:2007-03-25 发布日期:2007-03-25
  • 通讯作者: 徐虹

Cultiation and identification of mesenchymal stem cells derived from human umbilical cord blood and is clinical signification

  

  • Received:2006-03-17 Revised:2006-08-04 Online:2007-03-25 Published:2007-03-25

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 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,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,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。

Abstract: 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 sterilely with preservative-free heparin and the mononuclear cells were isolated from it, then plated the MNCs in 25-mm culture flasks containing DMEM/F12 medium. MSCs were obtained. 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.