书原代脐静脉内皮细胞与脐静脉内皮细胞株的论著培养方法及生物特性的比较古力热巴1聂永梅1*马依彤2杨毅宁2刘芬2陈邦党2吴江1，3(1新疆医科大学基础医学院生理学教研室新疆乌鲁木齐830011;2新疆心血管疾病重点实验室新疆乌鲁木齐830011;3四川大学华西基础医学与法医学院四川成都610041)【摘要】目的建立一套成功率高，细胞污染率少，获得较多的细胞数的人脐静脉内皮细胞(HUVEC)的体外分离方法，并且将原代脐静脉内皮细胞与EA．HY926人脐静脉内皮细胞株生物特性进行对比研究，为细胞培养及相关的医学基础研究提供依据。方法在新鲜(健康剖宫产产妇分娩后立即取新生胎儿脐带，长度15～20cm，取标本时间≤4h)脐静脉内注0．1%的Ⅱ型胶原酶消化获得内皮细胞，用M199生长液(含25%的内皮细胞生长因子)进行培养，EA．HY926细胞株用M199生长液。比较两组细胞的形态、超微结构、细胞纯度检测CD31抗体表达。结果①形态:原代HUVEC体外生长3～4d可铺满单层，细胞呈梭形、饱满，漩涡状排列生长，传代后可见管腔样结构;EA．HY926细胞株胞质多颗粒，2～3可铺满单层，可长成复层。②超微结构:透射电镜下，原代HUVEC里可发现较多的横切与纵切的韦伯潘力小体，在EA．HY926细胞株里较少。③细胞纯度检测:CD31抗体的表达率原代HUVEC细胞大于EA．HY926细胞株，两种细胞CD31表达差异有统计学意义(P﹤0．001)。结论脐静脉注Ⅱ型胶原酶消化法配合M199生长液可迅速获取大量内皮细胞，其生物学特性明显优于HUVEC株，而细胞株则以细胞数量多，培养方法简单，成活率高为特点，两者均是组织工程及相关医学基础研究较好的细胞来源。【关键词】原代人脐静脉内皮细胞EA．HY926细胞株细胞培养比较Comparationonculturemethodandbiologicalcharacteristicsbetweenprimaryhumanumbilicalveinendothelialcellsandhumanum-bilicalveinendothelialcellline．GULIRe－ba1，NIEYong－mei1*，MAYi－tong2，etal．1DepartmentofPhysiology，XinjiangMedicalUniver-sity，Urumqi，Xinjiang830011，China;2KeyLaboratoryofXinjiangCardiovascularDisease，Urumqi，Xinjiang830011，China．【Abstract】ObjectiveToestabilishaseparationmethodinvitro，withhighsuccessrateandlowcellcontaminationrate，anditisabletoobtainmorehumanumbilicalveinendothelialcells(HUVEC)andcomparewithEA．HY926humanumbilicalveinendothelialcelllineforstudyonbiologicalpropertiesandtoprovideaccordanceforcellcultureandrelatedbasicmedicalresearch．Methods0．1%solutionofcollagenasetypeⅡwasinjectedintofreshhumanumbilicalvein(obtainedfromumbilicalcordofhealthycesareansectionnewborn，withlengthof15－20cm，thetimeforobtainingspecimens≤4hours)togetendothelialcellsandcultivatedinM199growthmediumtoobtainEA．HY926celllineinM199growthmedium．Themorphologicalfeatures，ultrastructuralfindingsandexpressionofCD31antibodyfromcellswerecomparedbetweenthesetwogroupsofcells．Results⑴Cellmorphologicalexamination:theprimaryHUVECinvitrocultivatedfor3～4dcouldbecoveredwithasinglelay-er，theappearanceofcellswasspindleinshapewithfullandvortex－likearrangement，andlumen－likestructurecouldbefoundaftercellpas-sage．EA．HY926celllinehadmorecytoplasmicgranules，theycouldbecoveredwithsinglelayerafter2to3days，andtheycouldbegrowntomultiplelayers．⑵Ultrastructuralfeaturesofcells:undertransmissionelectronmicroscopy，primaryHUVECcouldbefoundinmorecross－sec-tionandlongitudinalsectionwithmoreweibel－paladebodies(WPB)，whereastheywerelessinEA．HY926celllines．⑶CellsexpressingCD31antibody:theexpressionofCD31antibodyinprimaryHUVECcells(85．36%)washigherthanthatofEA．HY926cellline(70．82%)．TtestforstatisticalanalysisbetweenthemwasP＜0．001，andthedifferenceinCD31expressionbetweenthesetwokindsofcellswasstatisticallysignifi-cant，anditwashigherinpri...
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