小鼠核移植胚胎干細胞系的建立及其全能性檢測.pdf

1、中國協和醫科大學中國醫學科學院碩士學位論文小鼠核移植胚胎干細胞系的建立及其全能性檢測姓名：張曉進申請學位級別：碩士專業：@指導教師：高紹榮郭巖20040901中國醫學科學院中國協和醫科大學北京生命科學研究所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馱comparedtoothermousestrains，1290G2FIgeneticbackgroundhasexhibitedadvantagesinntEScellderivationThesentEScell

7、sappearedtomaintainnormalESappearanceinvitroSixⅡtEScelllinesfrom1290G2FlmotileandsixBDFlntEScellslineswerechosentoinvestigatethepluripotencyOur髑uItsindicatedfiveofthetwelventEScelllinestestedexhibitedsimilarpluripotencyc

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