资源描述：
《汗腺细胞来源的诱导多能性干细胞构建及其鉴定》由会员上传分享，免费在线阅读，更多相关内容在工程资料-天天文库。
1、汗腺细胞来源的诱导多能性干细胞构建及其鉴定秦明德，梁含思(苏州大学江苏省干细胞与生物医用材料重点实验室，江苏苏州215000)摘要：木文中获得汗腺细胞来源的诱导多能干细胞(inducedpluripotentstemcell,iPS)。方法从手术废弃幼儿手指中提収出汗腺，体外培养，待汗腺细胞生长约10天后，以1x105的密度接种于6cm细胞培养血屮，向汗腺细胞培养基屮加入4种胚胎干细胞的转录因了基因:Oct4、Sox2、Klf4和c-Myc的慢病毒。2-3天后,消化接种到铺有滋养层细胞的6cm细胞培养皿屮，培养约10天后，挑选克隆和细胞形态与
2、胚胎干细胞和同的克隆，接种于铺有滋养层细胞的24孔板中。培养约10天后，再次挑选克隆和细胞形态与胚胎干细胞相同的克隆，接种于铺有滋养层细胞的6孔板中。RT-PCR和免疫荧光技术检测汗腺细胞标志基因：CEA、CK14和CK18；RT-PCR检测诱导后细胞的人胚胎干细胞标志基因:Oct4、Sox2>KLf4、c-Myc和nanong,免疫荧光技术检测是人胚胎干细胞表面标志基因：ssea3>ssea4、TRA-1-60和TRA-1-81以及核标志基因：Oct4o结果1、RT-PCR和免疫荧光技术检测从六指中提取的细胞表达汗腺细胞的标志基因。2、RT
3、-PCR和免疫荧光技术检测诱导后细胞表达人胚胎干细胞标志基因。结论汗腺细胞经过四因子的诱导后，可重编程为诱导多能干细胞。关键词：汗腺细胞；诱导多能干细胞；重编程中图分类号：Q254文献标志码：AConstructionandidentificationofSweatglandcellsderivedinducedpoluripotentstemcellsQINMingde,LIANGHansi(SoochowUniversity,JiangsuProvinceKeyLaboratoryofstemcellsandbiomatcrials,Ji
4、angSuSuZhou215000)Abstract:Inthispaper,inductionofpluripotentstemcellsfromsweatglandcells(inducedpluripotentstemcell,iPS).Methodssixchildrensweatgland,extractedfromtheculturedinvitro,thesweatglandcellsgrownforabout10days,with1*105wereinoculatedin6cmcellsinaPetridish,tosweat
5、glandcellcultureinto4humanembiyonicstemcelltranscriptionfactorgene:slowvirusOct4,Sox2,Klf4andc-Myc・After2-3days,digestedandinoculatedontohumantrophoblastcells6cmcellsinaPetridish,afterabout10daysofculture,theclonesandthemorphologyofthecellsandembryonicstemcellswereinoculate
6、dinthesame,witha24orificeplateoftrophoblastcells・Afterabout10daysoftraining,againselectedcloningandcellmorphologyandembryonicstemcellswereinoculatedinthesame,witha6orificeplateoftrophoblastcells・RT-PCRandimmunofluorescencedetectionofsweatglandcellmarkergenes:CEA,CK14andCK18
7、;cellmarkergeneofhumanembryonicstemcellswasdetectedbyRT-PCR:Oct4,Sox2,KLf4,c-MycandNanong,immunofluorescencedetectiontechnologyisthehumanembryonicstemcellsurfacemarkergenes:ssea3,ssea4,TRA-1・60andTRA・1・81aswellasanuclearmarkergene:Oct4.Resultsofthe1,RT-PCRandimmunofluoresce
8、ncedetectionfromsixreferstoextractcellmarkerexpressionofsweatglandcellgene・Humanem
