人脐带间充质干细胞的生物学特性及向神经样细胞分化的研究  

Biological characteristics of human umbilical cord mesenchymal stem cells and their differentiation into neural cells

作　　者：刘莎[1] 刘俊骞 郑彦茹 宫蕊[1] 初丽敏 魏树林[1] LIU Sha;LIU Junqian;ZHENG Yanru;GONG Rui;CHU Limin;WEI Shulin(The Third Hospital of Shijiazhuang City,Shijiazhuang 050011,China;The First Hospital of Hebei Medical University,Shijiazhuang 050031,China;The Emergency Center of Shijiazhuang City,Shijiazhuang 050000,China)

机构地区：[1]石家庄市第三医院,河北石家庄050011 [2]河北医科大学第一医院,河北石家庄050031 [3]石家庄市急救中心,河北石家庄050000

出　　处：《中国医药科学》2018年第5期37-39,176,共4页China Medicine And Pharmacy

基　　金：河北省医学科学研究重点课题(20150881)

摘　　要：目的探究人脐带间充质干细胞具有的生物学特性,并对其向神经样细胞的分化进行研究。方法检测人脐带间充质干细胞细胞表面的标记;以β巯基乙醇与丹参诱导其向神经细胞分化,并为分化细胞与未分化细胞进行免疫细胞化学鉴定。此外,半定量RT-PCR检测细胞神经相关基因表达。结果分离自人脐带的贴壁细胞在体外可以生长成类似成纤维细胞的形态,在未分化状态下可以维持稳定增殖形态,且体外增殖远超10代,这类细胞表面标记CD105、CD59、CD44、CD29有较高的表达,而造血细胞的表面标记CD117、CD45、CD34、CD33、CD28、CD14则无表达或低表达,而与其相同表达的还有移植免疫排斥相关的表面标记,如CD86、CD80、CD40。经诱导分化的细胞表达神经元标记β-Ⅲ类神经微管和神经微丝、神经干细胞标记巢蛋白、神经胶质细胞标记的胶质纤维酸性蛋白。经过RT-PCR检测可知,经丹参诱导后的间充质干细胞表达神经干细胞的相关基因Neatin,而诱导前后间充质干细胞全部表达神经细胞基因Pleiotrophin并且在诱导后表达增强。结论人脐带间充质干细胞具有易于培养扩增的生物学特性,其表达间充质干细胞的表面标记,低或不表达造血细胞、与移植排斥有关的细胞标记。同时,其可以分化为神经样细胞并表达神经细胞的基因与相关标记,这种细胞可以作为肝细胞的一个来源,用于中枢神经系统细胞的移植。Objective To investigate the biological characteristics of human umbilical cord mesenchymal stem cells(MSCs)and to study their differentiation into neuron-like cells.Methods The marker on the surface of human umbilical cord mesenchymal stem cells were labeled;The differentiate into neuronal cells were induced withβ-mercaptoethanol and salvia miltiorrhiza,and the immunocytochemical identification of the differentiated cells and undifferentiated cells was carried out.In addition,semi-quantitative RT-PCR was used to detect the expression of neuronal related genes.Results Adherent cells isolated from human umbilical cord could grow into fibroblast-like morphology in vitro and could maintain stable proliferative morphology in undifferentiated state.The proliferation of cells in vitro was well over 10 generations.The surface markers of these cells labeled CD105,CD59,CD44 and CD29 were higher expression,while the surface marker of hematopoietic cell labeled CD117,CD45,CD34,CD33,CD28,CD14 were no expression or low expression,and its expression in the same transplant rejection related surface markers such as CD86,CD80,CD40.The differentiated cells express neuronal markersβ-Ⅲneurotubules and neural microfilaments,neural stem cell marker nestin,and glial cell marker glial fibrillary acidic protein.RT-PCR detection results showed that mesenchymal stem cells induced by salvia miltiorrhiza express neatin,a gene related to neural stem cells,whereas all of the mesenchymal stem cells expressed neurotrophic factor Pleiotrophin before and after induction,and its expression was enhanced after induction.Conclusion Human umbilical cord mesenchymal stem cells have the biological characteristics of being easily cultured and expanded.They express the surface markers of mesenchymal stem cells,with or without expressing hematopoietic cells and cell markers related to graft rejection.At the same time,it can differentiate into neuron-like cells and express genes and related markers of nerve cells,which can be used as a source of hepatocy

关 键 词：人脐带 间充质干细胞 生物学特性 神经样细胞 分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]