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1、中南大学学报(医学版)2005,30(3)270JCentSouthUniv(MedSci)心肌细胞对骨髓间充质干细胞分化为心肌细胞的影响1,21,2,31,21,21,2王亭忠,马爱群,徐正云,蒋文慧,杜媛(西安交通大学1.第一医院心血管内科;2.医学院环境与疾病相关基因教育部重点实验室,西安710061)[摘要]目的:观察大鼠成体心肌细胞对骨髓间充质干细胞(mesenchymalstemcells,MSCs)定向分化的诱导作用。方法:应用Percoll’s密度梯度离心法分离培养大鼠MSCs,传2代后用流式细胞术检测其表面CD34,CD71和CD90的表达,然后用
2、Lipofectamine2000介导pEGFP2N3转染标记MSCs。将GFP标记的MSCs与新鲜分离的成体大鼠TM心肌细胞分别按接触、非接触(MILLICELL2HA半透膜分层培养)及心肌细胞条件培养液培养。1周后应用免疫荧光检测MSCsα2actin,desmin和cTnT的表达。结果:接触培养组中可见表达GFP的MSCs细胞同时表达α2actin,desmin和cTnT,而分层培养组及条件培养组中均未见α2actin,desmin和cTnT的表达。结论:大鼠成体心肌细胞与MSCs接触培养,可能诱导MSCs分化为心肌细胞。[关键词]骨髓间充质干细胞;成体心肌细
3、胞;诱导;分化[中图分类号]R329.2[文献标识码]A[文章编号]167227347(2005)0320270206Differentiationofmesenchymalstemcellsintocardiomyocytesinducedbycardiomyocytes1,21,2,31,21,21,2WANGTing2zhong,MAAi2qun,XUZheng2yun,JIANGWen2hui,DUYuan(1.DepartmentofCardiovascularMedicine,FirstHospital;2.KeyLaboratoryofEnvironm
4、entandGenesRelatedtoDiseases,Xi′anJiaotongUniversity,Xi′an710061,China)Abstract:ObjectiveToinvestigatetheroleofadultcardiomyocytesinthedifferentiationofmesenchymalstemcells(MSCs)intocardiomyocytes.MethodsRatMSCswereisolatedbyaPercoll’sgradientsolutionandculturedinlow2glucoseDulbecco’sm
5、odifiedEagle’smedium(DMEM).After2passages,cell2surfaceantigenCD34,CD71andCD90forratMSCsweredeterminedbyflowcytometry,andtheseMSCsweretransfectedwithpEGFP2N3byLipofectamine2000.ThenthoseMSCslabeledwithGFP,wereculturedincontacted,nocontactedandconditionedwithadultratmyocardiocytes.Immuno
6、fluo2rescencestainingagainstα2actin,desmin,andtroponin2Twereperformedafter1week.ResultsImmuno2fluorescencestainingwaspositiveagainstα2actin,desmin,andtroponin2TonMSCsincontactedculturegroup.Incontrast,noα2actin,desmin,andtroponin2TexpressiononMSCswereobservedinthenoncon2tactedculturegr
7、oupandtheconditionedculturegroup.ConclusionDirectcell2to2cellcontactbetweenMSCsandadultcardiomyocytesmayinducedifferentiationofMSCsintocardiomyocytes.Keywords:mesenchymalstemcells;adultcardiomyocytes;induce;differentiation[JCentSouthUniv(MedSci),2005,30(3):0270206][2～5]骨髓间充质干细胞(mesen
