小鼠核移植胚胎干細(xì)胞系的建立及其全能性檢測(cè).pdf

1、中國(guó)協(xié)和醫(yī)科大學(xué)中國(guó)醫(yī)學(xué)科學(xué)院碩士學(xué)位論文小鼠核移植胚胎干細(xì)胞系的建立及其全能性檢測(cè)姓名：張曉進(jìn)申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：@指導(dǎo)教師：高紹榮郭巖20040901中國(guó)醫(yī)學(xué)科學(xué)院中國(guó)協(xié)和醫(yī)科大學(xué)北京生命科學(xué)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