小鼠核移植胚胎干细胞系的建立及其全能性检测.pdf

1、中国协和医科大学中国医学科学院硕士学位论文小鼠核移植胚胎干细胞系的建立及其全能性检测姓名：张晓进申请学位级别：硕士专业：@指导教师：高绍荣郭岩20040901中国医学科学院中国协和医科大学北京生命科学研究所DerivationandpluripotencyanalysbofEScellsfrompreimplantationembryosproducedbysomaticcellnucleartransfer(sCNT)Abstract

2、Embryonicstem(ES)cellsarederivedfromtheinnercellmassofblastocystsproducedinvivoorbyinvitrofertilizationEScellsarepluripotentandcandifferentiateintoallthecelltypesofthreegermlayersRecentlyEScellshavebeenmcceesfullyderived

3、fromembryosproducedbysomaticcellnucleartransfer(SCNT)andithasbeenproposedimportantfortherapeuticpurposeandbasicresearchm‘thefutureInthepresentstudywederivedpluripotentembryonicstem(ES)cellsfrommurineblastocystsproducedby

4、somaticceilnucleartransfer(scNl3ThepluripotecyanddifferentiationpotentialofthentEScellsareinvestigatedbyRTPCRimmunocytochcmistrystainingandchimeraproductionWehavegeneratedmore70ntEScelllinesfromdifferentstrainofmicewhich

5、include1290G2F1andBDFImicethroughSCNTInthestudywefoundtheefficiencytosuccessfullyderiventEScellslinesfromclonedembryosvariedwithdifferentgeneticbackgroundofdonorcellsusedinSCNTTheBDFlmoⅡ卵ntESderivationefficiencyWas1062％o

6、G2(B6xCBA)n啦Swas10∞％，1290G2F1(129xB6xCBA)ntESWas3732％，B60G2F1(B6xB6xCBA)ⅡtESWas1016％andZEG(ICR)Wnsonly1％Obvious驮comparedtoothermousestrains，1290G2FIgeneticbackgroundhasexhibitedadvantagesinntEScellderivationThesentEScell

7、sappearedtomaintainnormalESappearanceinvitroSixⅡtEScelllinesfrom1290G2FlmotileandsixBDFlntEScellslineswerechosentoinvestigatethepluripotencyOur髑uItsindicatedfiveofthetwelventEScelllinestestedexhibitedsimilarpluripotencyc

8、omparedtonormalEScellsThegenesencodedtranscriptionfactorswhichareessentialformaintainingEScellspluripotencyincludingOct4，Sox2，NanogandRexI，wereexpressedintheⅡ倔ScellsastestedbyRTPCRandimmunocytoehemistrystainiⅡ昏Thediffere