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1、解剖科学进展ProgressofAnatomicalSciences2009,15(3):265~268星形胶质细胞条件培养液促进新生大鼠海马神经干细胞分化1,21111*王立芹,赵久红,唐源远,季丽莉,王振宇(1.中国医科大学基础医学院人体解剖学教研室,辽宁沈阳110001;2.山东聊城职业技术学院人体解剖学教研室,山东聊城252000)【摘要】目的探讨星形胶质细胞条件培养液(ACM)对神经干细胞(NSCs)体外分化的影响。方法行新生大鼠大脑皮质星形胶质细胞的纯化培养后收集ACM,将新生大鼠海马NSCs单克隆培养后行nestin免疫细胞荧光染色,诱导分化5d后行神经元特异性烯醇化酶(N
2、SE)、胶质纤维酸性蛋白(GFAP)、半乳糖脑苷脂(GALC)免疫细胞荧光染色;将单克隆培养的NSCs分为对照组和实验组,对照组为单纯NSCs培养液,实验组根据NSCs培养液与ACM比的不同分3组:A组(2:1),B组(1:1),C组(1:2)。各组培养1周,采用NSE免疫细胞荧光检测方法标记神经元并计数和统计学分析。结果纯化的星形胶质细胞胶质纤维酸性蛋白抗体标记阳性率为96.5%;单克隆培养的海马NSCs呈nestin阳性,诱导后呈NSE、GFAP和GALC阳性表达。ACM培养的海马NSCs各组分化为神经元的比例明显高于对照组(<0.01),其中A组的比例最高。结论ACM可以促进新生大
3、鼠海马NSCs向神经元分化。【关键词】星形胶质细胞条件培养液;神经干细胞;海马;新生大鼠【中图分类号】 Q189【文献标识码】A【文章编号】1006-2947(2009)03-0265-04Astrocyte-conditionedMediumPromotestheDifferentiationofNeuralStemCellsfromHippocampusofNeonateRats1,21111*WANGLi-qin,ZHAOJiu-hong,TANGYuan-yuan,JILi-li,WANGZhen-yu(1.DepartmentofHumanAnatomy,CollegeofBa
4、sicMedicine,ChinaMedicalUniversity,LiaoningShenyang110001;2.DepartmentofHumanAnatomy,LiaochengVocationalandTechnicalCollege,ShandongLiaocheng252000China)【Abstract】ObjectiveTostudytheinfluenceofastrocyte-conditionedmedium(ACM)onthedifferentiationofneuralstemcells(NSCs)invitro.MethodsAstrocytesfrom
5、thecerebralcortexofneonateratswereculturedandpurified,andACMwascollected.NSCsfromhippocampusofneonateratswereobtained,immunofluorescencestainwasusedtoshownestin-,neuronspecificenolase(NSE)-,glialfibrillaryacidicprotein(GFAP)-andgalactocerebroside(GALC)-positivecells.MonocloneculturedNSCsweredivid
6、edintocontrolgroup(NSCsmedium)andexperimentalgroups(withdifferentproportionsofNSCsmediumandACM:Agroup1:2,Bgroup1:1,Cgroup2:1).ThepercentageofNSEpositivecellswascountedandthedatawasanalyzedinstatistics.ResultsThepercentageofGFAPpositivecellswas96.5%inpurifiedastrocytes,theculturedNSCsshowednestinp
7、ositive,NSE,GFAPandGALCpositive.ThepercentageofNSEpositivecellsdifferentiatedfromNSCswasmuchhigherinexperimentalgroupsthanincontrolgroup(<0.01),withthehighestpercentageinAgroup.ConclusionACMcanpromotethedifferentiation