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1、中国组织工程研究第16卷第14期2012–04–01出版ChineseJournalofTissueEngineeringResearchApril1,2012Vol.16,No.14绵羊羊膜上皮细胞分离培养及鉴定*☆朱雪敏，李海军，米焱，杨银凤，关洪敏，曹贵方Isolation,cultureandidentiticationofOvisariesamnionepithelialcellsZhuXue-min,LiHai-jun,MiYan,YangYin-feng,GuanHong-min,CaoGui-f
2、angAbstractBACKGROUND:Theamnionepithelialcells(AECs)withstemcellspropertieshavebeensuccessfullyobtainedafterisolating,digestingandculturinghumanandratamnionandAECshavethepotentialtodifferentiateintocellsoftheendoderm(liver,lung,epithelium),mesoderm(bone,fat)
3、,andectoderm(neuralcells),OBJECTIVE:ToobtaintheOvisariesAECsanddetectthestemcellcharacteristics.METHODS:Ovisariesamniontissuewaspeeledoffbymechanicalwayandthelowvelocity-trypLE(pancreaticenzymereplacement)digestionmethodwasusedtoisolateandculturetheOvisaries
4、amnion,andtheOvisariesAECswereobtained.RESULTSANDCONCLUSION:TheimmunofluorescenceobservationshowedthatthelabeledproteinofAECssuchasOct-4,SSEA-1,SSEA-3,SSEA-4,TRA-1-60,TRA-1-81couldbeexpressedonOvisariesAECs.RT-PCRexperimentsconfirmedthatthegenesofOct-4,Sox-2
5、andRex-1wereexpressedandNanogwasnotexpressed.TheresultsindicatethattheOvisariesAECswithstemcellspropertieshavebeenachievedsuccessfully.ZhuXM,LiHJ,MiY,YangYF,GuanHM,CaoGF.Isolation,cultureandidentiticationofOvisariesamnionepithelialcells.ZhongguoZuzhiGongchen
6、gYanjiu.2012;16(14):2559-2562.[http://www.crter.cnhttp://en.zglckf.com]摘要背景：目前有研究发现人羊膜和大鼠羊膜分离消化培养后，可成功获得具有干细胞特性的人羊膜上皮细胞和鼠羊膜上皮细胞，在体外一些外源因子作用下具有向三胚层细胞分化的潜能。目的：获取绵羊羊膜上皮细胞，并对其干细胞特性进行鉴定。方法：采用机械方法剥离绵羊羊膜组织，运用低速旋转TrypLE(胰酶替代物)消化法对绵羊羊膜进行分离培养获得绵羊羊膜上皮细胞。结果与结论：细胞免疫荧光检测结
7、果表明绵羊羊膜上皮细胞表达Oct-4、SSEA-1、SSEA-3、SSEA-4、TRA-1-60、TRA-1-81等胚胎干细胞标记蛋白；RT-PCR技术鉴定结果显示绵羊羊膜上皮细胞的Oct-4、Sox-2、Rex-1全能性相关基因表达，而Nanog基因不表达。检测结果提示实验成功获得具有干细胞特性的绵羊羊膜上皮细胞。关键词：绵羊羊膜上皮细胞；分离；培养；细胞免疫荧光；RT-PCR缩略语注释：AECs：amnionepithelialcells，羊膜上皮细胞doi:10.3969/j.issn.1673-8225
8、.2012.14.018朱雪敏，李海军，米焱，杨银凤，关洪敏，曹贵方.绵羊羊膜上皮细胞分离培养鉴定[J].中国组织工程研究，2012，16(14):2559-2562.[http://www.crter.orghttp://cn.zglckf.com]LaboratoryofAnimalEmbryoandDevelopmentalBiology,InnerMongoliaAgricu
