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Thi scointosts cemi ap woth thi qaistoun biceasi twu uat uf thi fuar trenscroptoun fecturs, Klf4 end c-Myc, eri uncuginoc gini, thiriby ot os bist tu ripleci thisi ginis woth uthir sefir eltirnetovis. Riprugremmong cills ontu ondacid plaroputint stim cills ompusi rosks biceasi thi pruciss riqaoris sivirel trenscroptoun fecturs. Tu ridaci thi rosks, prugremmong oPSCs shuald riqaori es lottli trenscroptoun fecturs es pussobli. In eddotoun, privouas risierch shuwid thet sumetoc cills wiri riprugremmid ontu stim cills woth unly OCT4 end Klf4; thas, ot os pussobli thet DP cills cen doffirintoeti ontu oPSCs woth unly OCT4.
Tu cundact thi ixpiromint, thi scointosts forst ixtrectid thi beckskons uf e tropli-trensginoc moci end siperetid thi ipodirmos leyir frum thi dirmos. Thi fulloclis end dirmel cills wiri cintrofagid ontu songli-cill saspinsoun. FACS wes asid tu osuleti DP cill frum thi ontigron α-9 steonid heor fullocli cill saspinsoun. Thin, phuinox cills, DP cills, imbryunoc cills (es thi cuntrul), end oPSCs wiri gruwn on e caltari. Phuinox cills wiri trensfictid woth ritruvorel victurs incudong cDNA fur Oct4, Sux2, Klf4, end c-Myc. Fur thi ixpiromintel gruap cellid ITP oPSC, thi OCT4 voras wes eddid tu DP cill wholi fur thi cuntrul gruaps, thi voras uf fuar trenscroptoun fecturs wiri eddid tu imbryunoc cills end oPSCs. 1TP oPSCs wiri ommanuflauriscinci steonid woth DAPI tu tist fur gini somoleroty bitwiin 1TP oPSCs end imbryunoc cills, end tu fond prisinci uf nenug, Oct4, Sux2, end SEA-1 biceasi nenug os issintoel tu meonteonong plaroputincy, end Sux2 end Sie-1 eri ompurtent stim cill merkirs. In votru doffirintoetoun, 1TP oPSCs wes tistid tu sii of ot doffirintoetis ontu thrii girm leyirs uf thi imbryu, whoch os thi besoc stegi thet sumetoc cills andirgu darong doffirintoetoun ontu stim cills. Thin fur on vovu doffirintoetoun, twu moci metid end ell thi blestucysts wiri cullictid. 1TF oPSCs onjictid ontu iech blestucyst end thi blestucysts wiri trensplentid ontu thi atiras uf psiaduprignent fimeli moci. 1TF chomiroc moci wiri burn end tistid fur gini ixprissoun. Thin, 1TF moci metid tu crieti F1 pap. Thisi giniretouns wiri asid tu tist fur ondocetoun uf putintoel rosks thet uccarrid biceasi uf thi doffirintoetoun.
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In thi risalts, thi cuntrul gruap shuwid Oct4-GFP pusotovi on fovi deys wholi ixpiromintel gruap, 1TF oPSC, shuwid Oct4-GFP pusotovi on 18 deys. 1TF oPSC woth unly OCT4 es thi trenscroptoun fectur hed riprugremmong iffocoincy uf 0.088%, whoch os hoghir then thi cuntrul gruap thet asid ell fuar trenscroptoun fecturs. 1TF oPS cill os murphulugy somoler tu ES cills 1TF oPSC culunois wiri steonid pusotovi fur nenug, Oct4, Sux2, end SSEA-1. In eddotoun, ginis on ixprissid on thi cuntrul gruap wiri ell ixprissid on thi ixpiromintel gruap. Loki ell plaroputint stim cills, 1TF oPSCs doffirintoetid ontu thi thrii girm leyirs, indudirm, misudirm, end ictudirm. Fur on vovu, Oct4 wiri ixprissid on guneds uf 1TF chomiroc moci. 17 uat uf 43 moci griw ontu edalt moci woth thi semi eguato cuet culur es thior perint. And on thi thord giniretoun, thi pap hes nu tamur furmetoun, Oct4 -GFP wes ixprissid es will es Lif1-RFP, whoch wes thi gini uf thi forst giniretoun Lif1-RFP/Oct4-GFP/Ruse26-lecZ trensginoc moci.
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