<em>In Vitro</em> Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells

The placenta is the first organ to develop during embryogenesis and is required for the survival of the developing embryo. The placenta is comprised of ...

University of Pennsylvania

iPS cells can support full-term development of tetraploid blastocyst-complemented embryos.

Recent advances have demonstrated that the differentiated somatic cells could be reprogrammed into pluripotent state. Consequently, the reprogrammed somatic cells recapitulate the capacity to differentiate into specific cell lineages under appropriate culture conditions, which provides unlimited cell sources for cell transplantation-based therapy. In the present study, testicular Sertoli cells were successfully reprogrammed into pluripotent stem cells through somatic cell nuclear transfer (SCNT). Hematopoietic differentiation potential of the reprogrammed somatic cells was investigated in parallel to fertilization-derived ES (F-ES) cells. Our results demonstrated that the reprogrammed Sertoli cells (NT-ES) could efficiently differentiate into hematopoietic embryoid bodies (EBs). The hematopoietic-related genes including FLK-1, Bmp4, Runx1, etc. were dynamically expressed during the differentiation of the reprogrammed somatic cells in vitro. Transplantation of these differentiated reprogrammed cells into the bone marrow of irradiated mice could allow differentiation into different functional hematopoietic lineages in vivo. Moreover, blast-colony-forming cells (BL-CFCs) could be generated from both NT-ES and F-ES cells with similar efficiency in vitro. Our study indicates that the reprogrammed somatic cells possess the equivalent potency as F-ES cells in differentiating into functional hematopoietic cells.

Differentiation of reprogrammed somatic cells into functional hematopoietic cells.

&#x8868;&#x89C2;&#x9057;&#x4F20;&#x53D8;&#x5316;&#x7684;&#x52A8;&#x6001;&#x8C03;&#x63A7; DNA &#x7532;&#x57FA;&#x5316;&#x548C;&#x67D3;&#x8272;&#x8D28;&#x4FEE;&#x9970;&#x80DA;&#x80CE;&#x53D1;&#x80B2;&#x4E2D;&#x53D1;&#x6325;&#x5173;&#x952E;&#x4F5C;&#x7528;&#x3002;&#x867D;&#x7136;&#x6700;&#x8FD1;&#x7684;&#x7814;&#x7A76;&#x8868;&#x660E;&#xFF0C;&#x6838;&#x5FC3;&#x7EC4;&#x86CB;&#x767D;&#x7532;&#x57FA;&#x5316;&#x662F;&#x53EF;&#x9006;&#x7684;&#x4F46;&#x5F88;&#x5C11;&#x6709;&#x7814;&#x7A76;&#x8C03;&#x67E5;&#x4E86;&#x65B0;&#x53D1;&#x73B0;&#x7684;&#x7EC4;&#x86CB;&#x767D; demethylases &#x7684;&#x529F;&#x80FD;&#x5728;&#x80DA;&#x80CE;&#x53D1;&#x80B2;&#x8FC7;&#x7A0B;&#x3002;&#x5728;&#x672C;&#x7814;&#x7A76;&#x4E2D;&#xFF0C;&#x6211;&#x4EEC;&#x8C03;&#x67E5;&#x7684;&#x8868;&#x8FBE;&#x7279;&#x6027;&#x548C;&#x529F;&#x80FD;&#x7684; JMJD2C&#xFF0C;&#x5C5E;&#x4E8E;&#x542B;&#x6709; JmjC &#x57DF;&#x7684;&#x7EC4;&#x86CB;&#x767D; demethylases&#xFF0C;&#x9F20;&#x6807;&#x80DA;&#x80CE;&#x690D;&#x5165;&#x524D;&#x80DA;&#x80CE;&#x53D1;&#x80B2;&#x8FC7;&#x7A0B;&#x4E2D;&#x7684;&#x7EC4;&#x86CB;&#x767D;&#x8131;&#x3002;&#x6211;&#x4EEC;&#x53D1;&#x73B0; JMJD2C &#x9636;&#x6BB5;&#x5177;&#x4F53;&#x8868;&#x793A;&#x80DA;&#x80CE;&#x690D;&#x5165;&#x524D;&#x53D1;&#x5C55;&#x8FC7;&#x7A0B;&#x4E2D;&#xFF0C;&#x4E0E;&#x88AB;&#x89C2;&#x5BDF;&#x5230;&#x7684;&#x4E24;&#x4E2A;&#x5355;&#x5143;&#x683C;&#x4E2D;&#x5230; 8 &#x4E2A;&#x7EC6;&#x80DE;&#x9636;&#x6BB5;&#x6700;&#x9AD8;&#x7684;&#x6D3B;&#x52A8;&#x3002;JMJD2C &#x5728;&#x4E2D;&#x665A;&#x671F;&#x4E8C;&#x5375;&#x6BCD;&#x7EC6;&#x80DE;&#x5B64;&#x96CC;&#x6FC0;&#x6D3B;&#x8DDF;&#x67AF;&#x7AED;&#x5BFC;&#x81F4;&#x56CA;&#x80DA;&#x9636;&#x6BB5;&#x4E4B;&#x524D;&#x53D1;&#x5C55;&#x902E;&#x6355;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x7B26;&#x5408;&#x4EE5;&#x524D;&#x4F5C;&#x51FA;&#x7684;&#x7ED3;&#x8BBA; (ES) &#x80DA;&#x80CE;&#x5E72;&#x7EC6;&#x80DE;&#x4E2D;&#xFF0C;JMJD2C &#x7684;&#x67AF;&#x7AED;&#x5BFC;&#x81F4;&#x8BC4;&#x9009;&#x57FA;&#x56E0; Nanog &#x663E;&#x8457;&#x4E0B;&#x8C03;&#x80DA;&#x80CE;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x4E0E;&#x80DA;&#x80CE;&#x5E72;&#x7EC6;&#x80DE;&#x7814;&#x7A76;&#x4E0A;&#x4E00;&#x4EFD;&#x62A5;&#x544A;&#x6211;&#x4EEC;&#x89C2;&#x5BDF;&#x5230;&#x7684;&#x662F;&#x5176;&#x4ED6;&#x6F5C;&#x80FD;&#x7684;&#x57FA;&#x56E0;&#xFF0C;Pou5f1 &#x548C; Sox2&#x3001; &#x662F;&#x4E5F;&#x5927;&#x5E45;&#x4E0B;&#x8C03; JMJD2C &#x8017;&#x5C3D;&#x7684;&#x80DA;&#x80CE;&#x3002;&#x6B64;&#x5916;&#xFF0C;&#x5728;&#x65E9;&#x671F;&#x80DA;&#x80CE;&#x4E2D;&#x7684; JMJD2C &#x67AF;&#x7AED;&#x4E5F;&#x9020;&#x6210;&#x663E;&#x8457;&#x4E0B;&#x8C03; Myc &#x548C; Klf4 &#x57FA;&#x56E0;&#x4E0E;&#x7EC6;&#x80DE;&#x589E;&#x6B96;&#x76F8;&#x5173;&#x8054;&#x3002;&#x6211;&#x4EEC;&#x7684;&#x6570;&#x636E;&#x8868;&#x660E;&#x8FD9;&#x4E9B;&#x5173;&#x952E;&#x57FA;&#x56E0;&#x7684;&#x653E;&#x677E;&#x7BA1;&#x5236;&#x534F;&#x540C;&#x5BFC;&#x81F4;&#x89C2;&#x5BDF; JMJD2C &#x8017;&#x5C3D;&#x80DA;&#x80CE;&#x7684;&#x53D1;&#x80B2;&#x7F3A;&#x9677;&#x3002;

&#x7EC4;&#x86CB;&#x767D;&#x8131; JMJD2C &#x9636;&#x6BB5;&#x660E;&#x786E;&#x8868;&#x793A;&#xFF0C;&#x5C0F;&#x9F20;&#x80DA;&#x80CE;&#x690D;&#x5165;&#x524D;&#x80DA;&#x80CE;&#xFF0C;&#x5E76;&#x4E14;&#x662F;&#x80DA;&#x80CE;&#x53D1;&#x80B2;&#x7684;&#x5FC5;&#x8981;&#x6761;&#x4EF6;&#x3002;

Epigenetic modifications play a pivotal role in embryonic development by dynamically regulating DNA methylation and chromatin modifications. Although recent studies have shown that core histone methylation is reversible, very few studies have investigated the functions of the newly discovered histone demethylases during embryonic development. In the present study, we investigated the expression characteristics and function of JMJD2C, a histone demethylase that belongs to the JmjC-domain-containing histone demethylases, during preimplantation embryonic development of the mouse. We found that JMJD2C is stage-specifically expressed during preimplantation development, with the highest activity being observed from the two-cell to the eight-cell stage. Depletion of JMJD2C in metaphase II oocytes followed by parthenogenetic activation causes a developmental arrest before the blastocyst stage. Moreover, consistent with a previous finding in embryonic stem (ES) cells, depletion of JMJD2C causes a significant down-regulation of the pluripotency gene Nanog in embryos. However, contrary to a previous report in ES cells, we observed that other pluripotency genes, Pou5f1 and Sox2, are also significantly down-regulated in JMJD2C-depleted embryos. Furthermore, the depletion of JMJD2C in early embryos also caused significant down-regulation of the Myc and Klf4 genes, which are associated with cell proliferation. Our data suggest that the deregulation of these critical genes synergistically causes the developmental defects observed in JMJD2C-depleted embryos.

The histone demethylase JMJD2C is stage-specifically expressed in preimplantation mouse embryos and is required for embryonic development.

Modifications &eacute;pig&eacute;n&eacute;tiques jouent un r&ocirc;le central dans le d&eacute;veloppement embryonnaire en contr&ocirc;lant dynamiquement m&eacute;thylation de l&#39;ADN et des modifications de la chromatine. Bien que les &eacute;tudes r&eacute;centes ont montr&eacute; que la m&eacute;thylation histone core est r&eacute;versible, tr&egrave;s peu d&#39;&eacute;tudes ont &eacute;tudi&eacute; les fonctions de la d&eacute;m&eacute;thylases d&#39;histones nouvellement d&eacute;couvert durant le d&eacute;veloppement embryonnaire. Dans la pr&eacute;sente &eacute;tude, nous avons &eacute;tudi&eacute; les caract&eacute;ristiques de l&#39;expression et la fonction de JMJD2C, une histone d&eacute;m&eacute;thylase qui appartient &agrave; la d&eacute;m&eacute;thylases d&#39;histones contenant JmjC-domaine, au cours du d&eacute;veloppement embryonnaire pr&eacute;implantatoire de la souris. Nous avons trouv&eacute; que JMJD2C est &eacute;tape-sp&eacute;cifiquement exprim&eacute; au cours du d&eacute;veloppement pr&eacute;implantatoire, avec la plus forte activit&eacute; &eacute;tant observ&eacute;e entre les deux cellules et le stade de huit cellules. Appauvrissement de la couche de JMJD2C dans les ovocytes de m&eacute;taphase II suivie d&#39;activation parth&eacute;nog&eacute;n&eacute;tique provoque un arr&ecirc;t du d&eacute;veloppement avant le stade de blastocyste. En outre, conform&eacute;ment &agrave; une conclusion ant&eacute;rieure dans les cellules souches embryonnaires (ES), appauvrissement de la couche de JMJD2C provoque une r&eacute;gulation &agrave; la baisse importante du g&egrave;ne de la pluripotence Nanog chez les embryons. Cependant, contrairement &agrave; un pr&eacute;c&eacute;dent rapport, dans les cellules ES, nous avons observ&eacute; qu&#39;autres g&egrave;nes pluripotence, Pou5f1 et Sox2, sont &eacute;galement significativement diminu&eacute;es chez les embryons de JMJD2C appauvris. En outre, l&#39;&eacute;puisement des JMJD2C dans les jeunes embryons a &eacute;galement caus&eacute; r&eacute;gulation &agrave; la baisse significative des g&egrave;nes Myc et Klf4, qui sont associ&eacute;s &agrave; la prolif&eacute;ration cellulaire. Nos r&eacute;sultats sugg&egrave;rent que la d&eacute;r&eacute;glementation de ces g&egrave;nes critiques provoque en synergie les anomalies du d&eacute;veloppement observ&eacute;s chez les embryons de JMJD2C appauvris.

L&#39;histone d&eacute;m&eacute;thylase JMJD2C est exprim&eacute; en sc&egrave;ne-sp&eacute;cifiquement chez les embryons pr&eacute;implantatoires et est requis pour le d&eacute;veloppement embryonnaire.

Epigenetische Ver&auml;nderungen spielen eine zentrale Rolle in der embryonalen Entwicklung von dynamisch zu regulieren, DNA-Methylierung und Chromatin-Modifikationen. Obwohl neuere Studien, dass dieser Kern-Histon-Methylierung reversible ist gezeigt haben, haben nur sehr wenige Studien w&auml;hrend der Embryonalentwicklung die Funktionen der neu entdeckten Histon-Demethylasen untersucht. In der vorliegenden Studie untersuchten wir die Ausdruck-Merkmale und Funktion des JMJD2C, ein Histon-Demethylase, zu der der JmjC-Dom&auml;ne-haltigen Histon-Demethylasen w&auml;hrend der Pr&auml;implantationsdiagnostik Embryonalentwicklung der Maus geh&ouml;rt. Wir fanden, dass JMJD2C mit der h&ouml;chsten Aktivit&auml;t beobachtet aus der zwei-Zelle auf die acht-Zellen-B&uuml;hne B&uuml;hne-speziell w&auml;hrend der Pr&auml;implantationsdiagnostik Entwicklung ausgedr&uuml;ckt wird. Ersch&ouml;pfung der JMJD2C in Metaphase-II-Oozyten gefolgt von parthenogenetic Aktivierung bewirkt eine entwicklungsbedingte Verhaftung vor das Blastozystenstadium. Dar&uuml;ber hinaus f&uuml;hrt in &Uuml;bereinstimmung mit der fr&uuml;heren Feststellung in embryonale Stammzellen (ES), Ersch&ouml;pfung der JMJD2C eine erhebliche Herunterregulation der Pluripotenz gen Nanog Embryonen. Jedoch im Gegensatz zu einem vorherigen Bericht in ES-Zellen beobachteten wir, dass andere Pluripotenz Gene, Pou5f1 und Sox2, auch deutlich nach unten-in JMJD2C-abgereichertem Embryonen geregelt sind. Dar&uuml;ber hinaus verursacht die Ersch&ouml;pfung der JMJD2C im fr&uuml;hen Embryonen auch erhebliche Herunterregulation der Myc und Klf4 Gene, die Zellproliferation zugeordnet sind. Unsere Daten zeigen, dass die Deregulierung dieser kritischen Gene synergistisch bewirkt, die entwicklungsbedingte Defekte in JMJD2C-abgereichertem Embryonen beobachtet dass.

Die Histon-Demethylase JMJD2C ist B&uuml;hne-speziell in Pr&auml;implantationsdiagnostik Maus Embryonen ausgedr&uuml;ckt und ist erforderlich f&uuml;r die Embryonalentwicklung.

Modifiche epigenetiche giocano un ruolo fondamentale nello sviluppo embrionale regolando dinamicamente la metilazione del DNA e della cromatina modifiche. Anche se recenti studi hanno dimostrato che core la metilazione dell&#39;istone &egrave; reversibile, molto pochi studi hanno indagato le funzioni di demethylases l&#39;istone appena scoperti durante lo sviluppo embrionale. Nel presente studio, abbiamo studiato le caratteristiche di espressione e la funzione di JMJD2C, un demethylase istone che appartiene al demethylases dell&#39;istone JmjC-dominio-contenente, durante lo sviluppo embrionale preimpianto del mouse. Abbiamo trovato che JMJD2C fase-in particolare &egrave; espresso durante lo sviluppo preimpianto, con la pi&ugrave; alta attivit&agrave; osservato da due-cellula allo stadio di otto cellule. Deplezione di JMJD2C in ovociti II metafase seguita da attivazione nei provoca un arresto dello sviluppo prima allo stadio di blastocisti. Inoltre, coerente con le precedenti risultanze nelle cellule staminali embrionali (ES), deplezione di JMJD2C provoca una down-regulation significativo del gene della pluripotenza Nanog in embrioni. Tuttavia, contrariamente a una precedente relazione in cellule ES, abbiamo osservato che altri geni di pluripotenza, Pou5f1 e Sox2, sono anche significativamente gi&ugrave;-regolati in embrioni JMJD2C-esaurito. Inoltre, l&#39;esaurimento delle JMJD2C in embrioni in anticipo anche causato significativi down-regulation dei geni Myc e Klf4, che sono associati con la proliferazione cellulare. I nostri dati suggeriscono che la liberalizzazione di questi geni critici in sinergia causa i difetti di sviluppo osservati negli embrioni JMJD2C-esaurito.

L&#39;istone demethylase JMJD2C fase-in particolare &egrave; espresso in embrioni di topo preimpianto ed &egrave; necessaria per lo sviluppo embrionale.

&#x30A8;&#x30D4;&#x30B8;&#x30A7;&#x30CD;&#x30C6;&#x30A3;&#x30C3;&#x30AF;&#x4FEE;&#x98FE; DNA &#x306E;&#x30E1;&#x30C1;&#x30EB;&#x5316;&#x3068;&#x30AF;&#x30ED;&#x30DE;&#x30C1;&#x30F3;&#x4FEE;&#x98FE;&#x3092;&#x52D5;&#x7684;&#x306B;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x3053;&#x3068;&#x306B;&#x3088;&#x3063;&#x3066;&#x80DA;&#x306E;&#x6975;&#x3081;&#x3066;&#x91CD;&#x8981;&#x306A;&#x5F79;&#x5272;&#x3092;&#x518D;&#x751F;&#x3057;&#x307E;&#x3059;&#x3002;&#x6700;&#x8FD1;&#x306E;&#x7814;&#x7A76;&#x306F;&#x3001;&#x305D;&#x306E;&#x30B3;&#x30A2;&#x306E;&#x30D2;&#x30B9;&#x30C8;&#x30F3;&#x306E;&#x30E1;&#x30C1;&#x30EB;&#x5316;&#x306F;&#x4E0D;&#x53EF;&#x9006;&#x7684;&#x3067;&#x3042;&#x308B;&#x793A;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x304C;&#x3001;&#x975E;&#x5E38;&#x306B;&#x307B;&#x3068;&#x3093;&#x3069;&#x306E;&#x7814;&#x7A76;&#x306F;&#x840C;&#x82BD;&#x671F;&#x306E;&#x958B;&#x767A;&#x4E2D;&#x306B;&#x65B0;&#x305F;&#x306B;&#x767A;&#x898B;&#x3055;&#x308C;&#x305F;&#x30D2;&#x30B9;&#x30C8;&#x30F3;&#x30C7;&#x30E1;&#x30C1;&#x30E9;&#x30FC;&#x30BC;&#x306E;&#x6A5F;&#x80FD;&#x3092; &#x3057;&#x307E;&#x3059;&#x3002;&#x672C;&#x7814;&#x7A76;&#x3067;&#x306F;&#x3001;JMJD2C&#x3001;&#x30DE;&#x30A6;&#x30B9;&#x306E;&#x7740;&#x5E8A;&#x524D;&#x80DA;&#x767A;&#x751F;&#x4E2D;&#x306B;&#x5C5E;&#x3059;&#x308B; JmjC &#x30C9;&#x30E1;&#x30A4;&#x30F3;&#x542B;&#x6709;&#x30D2;&#x30B9;&#x30C8;&#x30F3;&#x30C7;&#x30E1;&#x30C1;&#x30E9;&#x30FC;&#x30BC;&#x306B;&#x30D2;&#x30B9;&#x30C8;&#x30F3;&#x8131;&#x30E1;&#x30C1;&#x30EB;&#x5316;&#x9175;&#x7D20;&#x306E;&#x6A5F;&#x80FD;&#x3068;&#x8868;&#x73FE;&#x7279;&#x5FB4;&#x3092;&#x691C;&#x8A0E;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x6211;&#x3005; &#x306F;&#x3001;JMJD2C &#x30B9;&#x30C6;&#x30FC;&#x30B8;&#x7279;&#x306B;&#x7740;&#x5E8A;&#x524D;&#x306E;&#x958B;&#x767A;&#x4E2D;&#x306F;&#x3001;2 &#x3064;&#x306E;&#x30BB;&#x30EB;&#x304B;&#x3089; 8 &#x7D30;&#x80DE;&#x671F;&#x306B;&#x89B3;&#x5BDF;&#x3055;&#x308C;&#x308B;&#x6700;&#x9AD8;&#x306E;&#x6D3B;&#x52D5;&#x3068;&#x8868;&#x73FE;&#x3055;&#x308C;&#x308B;&#x3053;&#x3068;&#x3092;&#x767A;&#x898B;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;JMJD2C &#x306E;&#x67AF;&#x6E07;&#x5358;&#x70BA;&#x6D3B;&#x6027;&#x5316;&#x306B;&#x3088;&#x3063;&#x3066;&#x5F93;&#x3044;&#x307E;&#x3057;&#x305F;&#x4E2D;&#x671F; II &#x5375;&#x6BCD;&#x7D30;&#x80DE;&#x306E;&#x767A;&#x9054;&#x306E;&#x902E;&#x6355;&#x524D;&#x306B;&#x80DA;&#x304C;&#x767A;&#x751F;&#x3057;&#x307E;&#x3059;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x4E00;&#x8CAB;&#x6027;&#x306E;&#x3042;&#x308B;&#x840C;&#x82BD;&#x671F;&#x306E;&#x830E; (ES) &#x7D30;&#x80DE;&#x306E;&#x524D;&#x3092;&#x898B;&#x3064;&#x3051;&#x308B;&#x3053;&#x3068;&#x306F;&#x3001;JMJD2C &#x306E;&#x67AF;&#x6E07;&#x591A;&#x80FD;&#x6027;&#x907A;&#x4F1D;&#x5B50; Nanog &#x306E;&#x5927;&#x5E45;&#x30C0;&#x30A6;&#x30F3;&#x898F;&#x5236;&#x80DA;&#x306E;&#x539F;&#x56E0;&#x3057;&#x307E;&#x3059;&#x3002;&#x3057;&#x304B;&#x3057;&#x306B;&#x53CD;&#x3057;&#x3066;&#x524D;&#x30EC;&#x30DD;&#x30FC;&#x30C8; ES &#x7D30;&#x80DE;&#x3067;&#x306F;&#x3001;&#x6211;&#x3005; &#x4ED6;&#x591A;&#x80FD;&#x6027;&#x907A;&#x4F1D;&#x5B50;&#x3001;Pou5f1 &#x3068; Sox2, &#x3082;&#x5927;&#x5E45;&#x306B;&#x30C0;&#x30A6;&#x30F3;-JMJD2C &#x67AF;&#x6E07;&#x80DA;&#x306E;&#x898F;&#x5236;&#x304C;&#x3055;&#x308C;&#x3066;&#x89B3;&#x5BDF;&#x3002;&#x3055;&#x3089;&#x306B;&#x3001;&#x521D;&#x671F;&#x80DA;&#x306B;&#x304A;&#x3051;&#x308B; JMJD2C &#x306E;&#x67AF;&#x6E07;&#x3082;&#x5927;&#x5E45;&#x30C0;&#x30A6;&#x30F3;&#x898F;&#x5236;&#x306E;&#x7D30;&#x80DE;&#x5897;&#x6B96;&#x306B;&#x95A2;&#x9023;&#x4ED8;&#x3051;&#x3089;&#x308C;&#x3066;&#x3044;&#x308B;&#x3001;Myc &#x304A;&#x3088;&#x3073; Klf4 &#x306E;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x539F;&#x56E0;&#x3067;&#x767A;&#x751F;&#x3002;&#x6211;&#x3005; &#x306E;&#x30C7;&#x30FC;&#x30BF;&#x306F;&#x3001;&#x3053;&#x308C;&#x3089;&#x91CD;&#x8981;&#x306A;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x898F;&#x5236;&#x7DE9;&#x548C;&#x304C;&#x76F8;&#x4E57;&#x7684; JMJD2C &#x67AF;&#x6E07;&#x80DA;&#x3067;&#x89B3;&#x6E2C;&#x3055;&#x308C;&#x305F;&#x7570;&#x5E38;&#x3092;&#x5F15;&#x304D;&#x8D77;&#x3053;&#x3059;&#x3053;&#x3068;&#x3092;&#x304A;&#x52E7;&#x3081;&#x3057;&#x307E;&#x3059;&#x3002;

&#x30D2;&#x30B9;&#x30C8;&#x30F3;&#x8131;&#x30E1;&#x30C1;&#x30EB;&#x5316;&#x9175;&#x7D20; JMJD2C &#x30B9;&#x30C6;&#x30FC;&#x30B8;&#x7279;&#x306B;&#x7740;&#x5E8A;&#x524D;&#x30DE;&#x30A6;&#x30B9;&#x80DA;&#x3067;&#x8868;&#x3055;&#x308C;&#x307E;&#x3059;&#x3001;&#x840C;&#x82BD;&#x671F;&#x306E;&#x958B;&#x767A;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3059;&#x3002;

Epigenetic &#xC218;&#xC815; DNA &#xBA54; &#xD2F8; &#xD654;&#xC640; chromatin &#xC218;&#xC815; &#xB3D9;&#xC801;&#xC73C;&#xB85C; &#xC870;&#xC808; &#xD558; &#xC5EC; &#xBC30;&#xC544; &#xAC1C;&#xBC1C;&#xC5D0; &#xC911;&#xCD94;&#xC801;&#xC778; &#xC5ED;&#xD560;&#xC744; &#xB2F4;&#xB2F9;&#xD560;. &#xCD5C;&#xADFC; &#xC5F0;&#xAD6C;&#xB294; &#xCF54;&#xC5B4; &#xD788;&#xC2A4;&#xD1A4; &#xBA54; &#xD2F8; &#xD654;&#xB294; &#xAC00;&#xC5ED; &#xAC83;&#xC73C;&#xB85C; &#xB098;&#xD0C0;&#xB0AC;&#xC2B5;&#xB2C8;&#xB2E4;, &#xD558;&#xC9C0;&#xB9CC; &#xC544;&#xC8FC; &#xC18C;&#xC218;&#xC758; &#xC5F0;&#xAD6C; &#xBC30;&#xC544; &#xAC1C;&#xBC1C; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xC0C8;&#xB85C; &#xBC1C;&#xACAC;&#xB41C; &#xB41C; &#xD788;&#xC2A4;&#xD1A4; demethylases &#xAE30;&#xB2A5; &#xC870;&#xC0AC;. &#xD604;&#xC7AC;&#xC758; &#xC5F0;&#xAD6C;&#xC5D0; &#xC6B0;&#xB9AC; &#xC2DD; &#xD2B9;&#xC131;&#xACFC; JMJD2C, &#xB9C8;&#xC6B0;&#xC2A4;&#xC758; preimplantation &#xBC30;&#xC544; &#xAC1C;&#xBC1C; &#xD558;&#xB294; &#xB3D9;&#xC548; JmjC &#xB3C4;&#xBA54;&#xC778; &#xD3EC;&#xD568; &#xB41C; &#xD788;&#xC2A4;&#xD1A4; demethylases&#xC5D0; &#xC18D;&#xD558;&#xB294; &#xD788;&#xC2A4;&#xD1A4; demethylase&#xC758; &#xAE30;&#xB2A5;&#xC744; &#xC870;&#xC0AC;. &#xC6B0;&#xB9AC;&#xB294; JMJD2C 8 &#xC140; &#xB2E8;&#xACC4;&#xB85C; 2 &#xC140;&#xC5D0;&#xC11C; &#xAD00;&#xCC30; &#xB418; &#xACE0; &#xAC00;&#xC7A5; &#xB192;&#xC740; &#xD65C;&#xB3D9; preimplantation &#xAC1C;&#xBC1C; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xBB34;&#xB300;&#xC5D0; &#xAD6C;&#xCCB4;&#xC801;&#xC73C;&#xB85C; &#xD45C;&#xC2DC; &#xB429;&#xB2C8;&#xB2E4; &#xBC1C;&#xACAC;. JMJD2C &#xBD84;&#xC5F4; II oocytes parthenogenetic &#xD65C;&#xC131;&#xD654; &#xB4A4;&#xC5D0; &#xACE0;&#xAC08; blastocyst &#xBB34;&#xB300; &#xC804;&#xC5D0; &#xBC1C;&#xB2EC; &#xB9C8;&#xBE44;&#xB97C; &#xBC1C;&#xC0DD; &#xD569;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, &#xBC30;&#xC544; &#xC904;&#xAE30; (ES&#xC758;) &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xC774;&#xC804; &#xCC3E;&#xAE30;&#xC640; &#xC77C;&#xCE58;, JMJD2C &#xACE0;&#xAC08; &#xD558;&#xBA74; &#xC0C1;&#xB2F9;&#xD55C; &#xB2E4;&#xC6B4; &#xADDC;&#xC81C; pluripotency &#xC720;&#xC804;&#xC790; Nanog &#xBC30;&#xC544;&#xC5D0;&#xC11C;. &#xADF8;&#xB7EC;&#xB098;, ES &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xC774;&#xC804; &#xBCF4;&#xACE0;&#xC11C;&#xC5D0; &#xBC18;&#xB300;&#xB294; &#xB2E4;&#xB978; pluripotency &#xC720;&#xC804;&#xC790; Pou5f1 &#xBC0F; Sox2, &#xD06C;&#xAC8C; &#xB2E4;&#xC6B4;-&#xADDC;&#xC81C; JMJD2C &#xACE0;&#xAC08; &#xBC30;&#xC544;&#xC5D0;&#xC11C; &#xAD00;&#xCC30; &#xD569;&#xB2C8;&#xB2E4;. &#xB610;&#xD55C;, &#xCD08;&#xAE30; &#xBC30;&#xC544;&#xC5D0; JMJD2C &#xACE0;&#xAC08;&#xB3C4; &#xC0C1;&#xB2F9;&#xD55C; &#xB2E4;&#xC6B4; &#xB808; &#xADE4; &#xB808;&#xC774; &#xC158; &#xAD00;&#xB828; &#xC138;&#xD3EC; &#xC99D;&#xC2DD; &#xB41C; Myc &#xBC0F; Klf4 &#xC720;&#xC804;&#xC790;&#xC758; &#xBC1C;&#xC0DD; &#xD569;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xC758; &#xB370;&#xC774;&#xD130; &#xC911;&#xC694; &#xD55C; &#xC720;&#xC804;&#xC790;&#xC758; &#xADDC;&#xC81C; &#xC644;&#xD654; synergistically &#xBC1C;&#xB2EC; &#xACB0;&#xD568; JMJD2C &#xACE0;&#xAC08; &#xBC30;&#xC544;&#xC5D0;&#xC11C; &#xAD00;&#xCC30; &#xD558;&#xBA74; &#xAC83;&#xC774; &#xC88B;&#xC2B5;&#xB2C8;&#xB2E4;.

&#xD788;&#xC2A4;&#xD1A4; demethylase JMJD2C preimplantation &#xB9C8;&#xC6B0;&#xC2A4; &#xBC30;&#xC544;&#xC5D0;&#xC11C; &#xB2E8;&#xACC4; &#xAD6C;&#xCCB4;&#xC801;&#xC73C;&#xB85C; &#xD45C;&#xD604; &#xB418; &#xACE0; &#xBC30;&#xC544; &#xBC1C;&#xB2EC;&#xC5D0; &#xB300; &#xD55C; &#xD544;&#xC694; &#xD569;&#xB2C8;&#xB2E4;.

Las modificaciones epigen&eacute;ticas juegan un papel fundamental en el desarrollo embrionario regulando din&aacute;micamente la metilaci&oacute;n del ADN y cromatina modificaciones. Aunque estudios recientes han demostrado que metilaci&oacute;n de histonas de base es reversible, muy pocos estudios han investigado las funciones de la histona reci&eacute;n descubierto demetilasas durante el desarrollo embrionario. En este estudio, investigamos las caracter&iacute;sticas de la expresi&oacute;n y funci&oacute;n de JMJD2C, un demetilasa histona que pertenece a la que contiene JmjC y dominio histona demetilasas, durante el desarrollo embrionario preimplantacional del rat&oacute;n. Encontramos que JMJD2C etapa espec&iacute;fica se expresa durante el desarrollo Preimplantacional, con la mayor actividad se observa de las dos c&eacute;lulas a la etapa de ocho c&eacute;lulas. Agotamiento de JMJD2C en ovocitos II metafase seguido partenogen&eacute;tica activaci&oacute;n provoca una detenci&oacute;n del desarrollo antes de la etapa de blastocisto. Por otra parte, consistente con un anterior hallazgo en c&eacute;lulas madre embrionarias (ES), agotamiento de JMJD2C causa una importante regulaci&oacute;n del gen pluripotencia Nanog en embriones. Sin embargo, al contrario de un informe anterior en c&eacute;lulas madre embrionarias, observamos que otros genes de pluripotencia, Pou5f1 y Sox2, son tambi&eacute;n significativamente regula en embriones JMJD2C-agotado. Adem&aacute;s, el agotamiento de los JMJD2C en embriones tempranos tambi&eacute;n caus&oacute; importante abajo-regulaci&oacute;n de los genes Myc y Klf4, que se asocian a la proliferaci&oacute;n celular. Nuestros datos sugieren que la desregulaci&oacute;n de estos genes cr&iacute;ticos causa sin&eacute;rgicamente los defectos de desarrollo observados en embriones JMJD2C-agotado.

La histona demetilasa JMJD2C etapa espec&iacute;fica se expresa en embriones de rat&oacute;n preimplantaci&oacute;n y se requiere para el desarrollo embrionario.

Androgenetic embryonic stem (aES) cells, produced by pronuclear transplantation, offer an important autologous pluripotent stem cell source. However, the isolation of aES cells, particularly individual-specific aES cells, with the use of fertilized embryos has limited the practical applications of this technology in humans. In this study, we applied a new approach, essentially described as somatic cell nuclear transfer, and generated three aES cell line types with the use of spermatogenic cells including primary spermatocytes, round spermatids, and mature spermatozoa as donor cells, omitting the need to use fertilized embryos. Although abnormality of chimeras and absent germline competency indicated that all three types of aES cells exhibited limited pluripotency, the epigenetic status of the aES cell lines tended to resemble normal ES cells during long-term culture, and some parental-specific imprinted genes were expressed at levels comparable to those of normal ES cells. Furthermore, the histocompatibility of the aES cells was investigated by transplanting the differentiation progenies of the aES cells into major histocompatibility (MHC)-matched and -mismatched recipient mice. The results indicated that these aES cells were histocompatible with MHC-matched mice after transplantation. Our study provides evidence that MHC-competent autologous aES cells could be generated from different spermatogenic cells using nuclear transfer into oocytes, a process that could avoid the use of fertilized embryos.

Generation of histocompatible androgenetic embryonic stem cells using spermatogenic cells.

Differentiated somatic cells of various species can be reprogrammed into induced pluripotent stem cells (iPSCs) by ectopically expressing a combination of several transcription factors that are highly enriched in embryonic stem cells (ESCs). The generation of iPSCs in large animals has raised the possibility of producing genetically modified large animals through the nuclear transplantation approach. However, it remains unknown whether iPSCs could be used for generating cloned animals through the nuclear transfer method. Here, we show the successful production of viable cloned mice from inducible iPSCs through the nuclear transfer approach, and the efficiency is similar to that of using ESCs derived via normal fertilization. Furthermore, the cloned mice are fertile and can produce second-generation offspring. These efforts strengthen the possibility of utilizing iPSCs to generate gene-modified large animals for pharmaceutical purposes in the future.

Mice cloned from induced pluripotent stem cells (iPSCs).

It has been recently reported that the regulatory circuitry formed by OCT4, miR-302, and NR2F2 controls both pluripotency and neural differentiation of human embryonic stem cells (hESCs). We show here that JMJD1C, a histone 3 lysine 9 (H3K9) demethylase expressed in hESCs, directly interacts with this circuitry. hESCs with stable knockdown of JMJD1C remain pluripotent while having reduced miR-302 expression, decreased BMP signaling, and enhanced TGFβ signaling. JMJD1C binds to the miR-302 promoter and reduces H3K9 methylation. Withdrawal of basic fibroblast growth factor (bFGF) from the culture induces neural differentiation of the knockdown, but not the control, cells within 3 days, accompanied by elevated NR2F2 expression. This can be attenuated with miR-302 mimics or an H3K9 methytransferase inhibitor. Together, our findings suggest that JMJD1C represses neural differentiation of hESCs at least partially by epigenetically sustaining miR-302 expression and that JMJD1C knockdown is sufficient to trigger neural differentiation upon withdrawal of exogenous bFGF.

Epigenetic regulation of miR-302 by JMJD1C inhibits neural differentiation of human embryonic stem cells.

Polycomb group proteins mediate transcriptional silencing in diverse developmental processes. Sex chromosomes undergo chromosome-wide transcription silencing during male meiosis. Here we report that mouse SCML2 (Sex comb on midleg-like 2), an X chromosome-encoded polycomb protein, is specifically expressed in germ cells, including spermatogonia, spermatocytes, and round spermatids. SCML2 associates with phosphorylated H2AX and localizes to the XY body in spermatocytes. Loss of SCML2 in mice causes defective spermatogenesis, resulting in sharply reduced sperm production. SCML2 interacts with and recruits a deubiquitinase, USP7, to the XY body in spermatocytes. In the absence of SCML2, USP7 fails to accumulate on the XY body, whereas H2A monoubiquitination is dramatically augmented in the XY chromatin. Our results demonstrate that the SCML2/USP7 complex constitutes a novel molecular pathway in modulating the epigenetic state of sex chromosomes during male meiosis.

Polycomb protein SCML2 associates with USP7 and counteracts histone H2A ubiquitination in the XY chromatin during male meiosis.

Females have a greater immunological advantage than men, yet they are more prone to autoimmune disorders. The basis for this sex bias lies in the X chromosome, which contains many immunity-related genes. Female mammals use X chromosome inactivation (XCI) to generate a transcriptionally silent inactive X chromosome (Xi) enriched with heterochromatic modifications and XIST/Xist RNA, which equalizes gene expression between the sexes. Here, we examine the maintenance of XCI in lymphocytes from females in mice and humans. Strikingly, we find that mature naïve T and B cells have dispersed patterns of XIST/Xist RNA, and they lack the typical heterochromatic modifications of the Xi. In vitro activation of lymphocytes triggers the return of XIST/Xist RNA transcripts and some chromatin marks (H3K27me3, ubiquitin-H2A) to the Xi. Single-cell RNA FISH analysis of female T cells revealed that the X-linked immunity genes CD40LG and CXCR3 are biallelically expressed in some cells. Using knockout and knockdown approaches, we find that Xist RNA-binding proteins, YY1 and hnRNPU, are critical for recruitment of XIST/Xist RNA back to the Xi. Furthermore, we examined B cells from patients with systemic lupus erythematosus, an autoimmune disorder with a strong female bias, and observed different XIST RNA localization patterns, evidence of biallelic expression of immunity-related genes, and increased transcription of these genes. We propose that the Xi in female lymphocytes is predisposed to become partially reactivated and to overexpress immunity-related genes, providing the first mechanistic evidence to our knowledge for the enhanced immunity of females and their increased susceptibility for autoimmunity.

Unusual maintenance of X chromosome inactivation predisposes female lymphocytes for increased expression from the inactive X.

Long noncoding RNAs (lncRNAs) can regulate gene expression in a cell-specific fashion during development. Here, we identify a novel lncRNA from the X chromosome that we named lncRHOXF1 and which is abundantly expressed in trophectoderm and primitive endoderm cells of human blastocyst-stage embryos. lncRHOXF1 is a spliced and polyadenylated lncRNA about 1 kb in length that is found in both the nuclear and cytoplasmic compartments of in vitro differentiated human trophectoderm progenitor cells. Gain-of-function experiments in human embryonic stem cells, which normally lack lncRHOXF1 RNA, revealed that lncRHOXF1 reduced proliferation and favored cellular differentiation. lncRHOXF1 knockdown using small interfering RNAs (siRNAs) in human trophectoderm progenitors increased expression of viral response genes, including type I interferon. Sendai virus infection of human trophectoderm progenitor cells increased lncRHOXF1 RNA levels, and siRNA-mediated disruption of lncRHOXF1 during infection reduced the expression of viral response genes leading to higher virus replication. Thus, lncRHOXF1 RNA is the first example of a lncRNA that regulates the host response to viral infections in human placental progenitor cells, and we propose that it functions as a repressor of the viral response during early human development.

lncRHOXF1, a Long Noncoding RNA from the X Chromosome That Suppresses Viral Response Genes during Development of the Early Human Placenta.

It has been suggested that excessive apoptosis in intervertebral disc cells induced by inflammatory cytokines, such as interleukin (IL)-1β, is related to the process of intervertebral disc degeneration (IVDD). Hydrogen sulfide (H2S), a gaseous signaling molecule, has drawn attention for its anti-apoptosis role in various pathophysiological processes in degenerative diseases. To date, there has been no investigation of the correlation of H2S production and IVDD or of the effects of H2S on IL-1β-induced apoptosis in nucleus pulposus (NP) cells. Here, we found that the expression levels of cystathionine β-synthase (CBS) and cystathionine γ-lyase (CSE), two key enzymes in the generation of H2S, were significantly decreased in human degenerate NP tissues as well as in IL-1β-treated NP cells. NaHS (H2S donor) administration showed a protective effect by inhibiting the endoplasmic reticulum (ER) stress response and mitochondrial dysfunction induced by IL-1β stimulation in vitro, the effect was related to activation of the PI3K/Akt and ERK1/2 signaling pathways. Suppression of these pathways by specific inhibitors, LY294002 and PD98059, partially reduced the protective effect of NaHS. Moreover, in the percutaneous needle puncture disc degeneration rat tail model, disc degeneration was partially reversed by NaHS administration. Taken together, our results suggest that H2S plays a protective role in IVDD and the underlying mechanism involves PI3K/Akt and ERK1/2 signaling pathways-mediated suppression of ER stress and mitochondrial dysfunction in IL-1β-induced NP cells.

Hydrogen sulfide protects against endoplasmic reticulum stress and mitochondrial injury in nucleus pulposus cells and ameliorates intervertebral disc degeneration.

Jianle Wang

Department of Biomedical Sciences,
School of Veterinary Medicine,
Department of Biomedical Sciences, School of Veterinary Medicine

University of Pennsylvania

In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells

Jianle Wang

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Biomedical Sciences,School of Veterinary Medicine,Department of Biomedical Sciences, School of Veterinary Medicine

University of Pennsylvania

In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells

Department of Biomedical Sciences,
School of Veterinary Medicine,
Department of Biomedical Sciences, School of Veterinary Medicine