nucleofection-and-in-vivo-propagation-of-chicken-eimeria-parasites

Nucleofection and In Vivo Propagation of Chicken <em>Eimeria</em> Parasites

Transfection is a technical process through which genetic material, such as DNA and double-stranded RNA, are delivered into cells to modify the gene of ...

Peking University

Severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV) is the principal etiologic agent of SARS. We analyzed serum samples obtained from 623 patients with SARS in Beijing, to determine whether infection with SARS-CoV can elicit neutralizing antibodies (NAbs).

Neutralizing antibodies in patients with severe acute respiratory syndrome-associated coronavirus infection.

To identify whether Nestin-positve cells or Nestin-negative cells in pancreas enrich potential pancreatic stem/progenitor cells.

NestinnegCD24low/- population from fetal Nestin-EGFP transgenic mice enriches the pancreatic endocrine progenitor cells.

Whether bone marrow (BM) hematopoietic stem/progenitor cells can directly differentiate into nonhematopoietic cells remains controversial. The aim of this study is to further investigate the potentiality of BM hematopoietic progenitor cells to convert into hepatocytes in vitro.

Failure of hepatocyte marker-expressing hematopoietic progenitor cells to efficiently convert into hepatocytes in vitro.

Human embryonic stem cells (hES cells) have unlimited self-renewal capacity and can differentiate into most, if not all, possible cell types. This unique property makes them valuable not only for investigation of early developmental processes, but also for regenerative medicine. Mesoderm-derived cardiac cells and hematopoietic cells both have the potential for various therapeutic applications. However, efficient induction of hES cell differentiation into mesoderm remains a challenge. Here, we showed that treatment of hES cells with bone morphogenetic protein-4 (BMP-4) exhibited differential effects: long-term treatment results in trophoblast and extra-embryonic endoderm differentiation, whereas short-term treatment can promote early mesoderm induction. The induction of mesoderm in hES cells occurs at a high efficiency as measured using several markers, such as Brachyury, WNT3, and MIXL1 expression. Moreover, these mesoderm progenitor cells can differentiate into cardiac and hematopoietic lineages in vitro. Further analysis showed that the mesoderm-inducing capacity of BMP-4 requires endogenous FGF and TGF-beta/Nodal/activin signaling activities. Thus, our results uncover a novel role for BMP-4 in regulation of hES cell differentiation and should provide insights into the mechanism of mesoderm induction in hES cells.

Short-term BMP-4 treatment initiates mesoderm induction in human embryonic stem cells.

The G-protein-coupled receptor 56 (GPR56) plays important roles in brain development and tumorigenesis. cDNA data suggest that GPR56 has potential to become a neural stem cell (NSC) or neural progenitor cell (NPC) marker. However, expression of GPR56 protein in human NSC/NPCs was not explored. Using specific antibodies and immunochemistry, we showed that GPR56 was highly expressed in nestin-positive NSC/NPCs in the ventricular/subventricular zone of human and mouse fetal brains, and in cultured neurospheres derived from both human and mouse fetal brains. Downregulation of GPR56 protein occurred earlier than that of nestin in differentiating neurosphere cultures. Loss of GPR56 protein was also evident in well-differentiated glial fibrillary acidic protein-positive astrocytes and betaIII-tubulin-positive neurons. Our data suggest that GPR56 can be used as an NSC/NPC marker within the neural cell lineage, especially in combination with nestin.

GPR56 is highly expressed in neural stem cells but downregulated during differentiation.

We show that 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2) induces apoptosis and down-regulates the expression of enoyl-CoA hydratase short chain 1 (ECHS1) and peroxiredoxin 3 (PRDX3) in human breast cancer MCF-7 cells. The decrease of ECHS1 and PRDX3 was validated by Western blot and quantitative real-time reverse transcription-PCR in MCF-7 and other carcinoma cells. Knockdown and over-expression of ECHS1 and/or PRDX3 further supported the key role of ECHS1 and PRDX3 in regulation of PP2-induced apoptosis. These results suggest a possible apoptotic pathway whereby down-regulation of ECHS1 and PRDX3 potentiates PP2-induced apoptosis in MCF-7 cells.

The role of enoyl-CoA hydratase short chain 1 and peroxiredoxin 3 in PP2-induced apoptosis in human breast cancer MCF-7 cells.

&#x80DA;&#x80CE;&#x9020;&#x8840;&#x529F;&#x80FD;&#x662F;&#x4E00;&#x4E2A;&#x590D;&#x6742;&#x7684;&#x8FC7;&#x7A0B;&#x3002;&#x9610;&#x53D1;&#x4ECE;&#x80DA;&#x80CE;&#x5E72;&#x7EC6;&#x80DE;&#x7684;&#x9020;&#x8840;&#x5206;&#x5316;&#x8C03;&#x63A7;&#x7684;&#x673A;&#x5236;&#x5C06;&#x4F7F;&#x6211;&#x4EEC;&#x80FD;&#x591F;&#x5EFA;&#x7ACB;&#x4E00;&#x9879;&#x6218;&#x7565;&#xFF0C;&#x6709;&#x6548;&#x5730;&#x751F;&#x6210;&#x9020;&#x8840;&#x7EC6;&#x80DE;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x89C4;&#x7BA1;&#x4ECE;&#x4EBA;&#x7C7B;&#x80DA;&#x80CE;&#x5E72;&#x7EC6;&#x80DE; (hESCs) &#x9020;&#x8840;&#x7956;&#x7EC6;&#x80DE;&#x7684;&#x751F;&#x6210;&#x673A;&#x5236;&#x4ECD;&#x7136;&#x662F;&#x672A;&#x77E5;&#x7684;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#xFF0C;&#x51FA;&#x73B0;&#x4E86; CD43(+) &#x4ECE;&#x9020;&#x8840;&#x9020;&#x8840;&#x7EC6;&#x80DE;&#x7684;&#x57FA;&#x7840;&#x4E0A; &#xFF08;&#x4ED6;&#xFF09; &#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#xFF0C;&#x6211;&#x4EEC;&#x8868;&#x660E; VEGF &#x662F;&#x5FC5;&#x8981;&#x548C;&#x8DB3;&#x591F;&#x7684;&#x548C;&#x8FD9; bFGF &#x662F;&#x534F;&#x540C;&#x4E0E; VEGF &#x6307;&#x5B9A;&#x4ED6;&#x7684;&#x5355;&#x5143;&#x683C;&#x5E76;&#x968F;&#x540E;&#x8FC7;&#x6E21;&#x5230; CD43(+) &#x9020;&#x8840;&#x7EC6;&#x80DE;&#x3002;&#x91CD;&#x8981;&#x7684;&#x662F;&#xFF0C;&#x6211;&#x4EEC;&#x8BA4;&#x5B9A; TGF&beta; &#x65B0;&#x578B;&#x4FE1;&#x53F7;&#x8C03;&#x8282;&#x9020;&#x8840;&#x7EC6;&#x80DE;&#x53D1;&#x80B2;&#xFF0C;&#x50CF; TGF&beta; &#x6291;&#x5236;&#x5242; SB 431542 &#x5927;&#x5927;&#x63D0;&#x5021;&#x4ECE;&#x4ED6;&#x7684;&#x8FC7;&#x6E21;&#x671F;&#x95F4; hESC &#x5206;&#x5316;&#x7EC6;&#x80DE; CD43(+) &#x9020;&#x8840;&#x7956;&#x7EC6;&#x80DE; (HPCs)&#x3002;&#x901A;&#x8FC7;&#x5728;&#x9020;&#x8840;&#x5206;&#x5316;&#x8FC7;&#x7A0B;&#x5B9A;&#x4E49;&#x8FD9;&#x4E9B;&#x4FE1;&#x53F7;&#x7684;&#x5173;&#x952E;&#x56E0;&#x7D20;&#xFF0C;&#x6211;&#x4EEC;&#x6709;&#x6548;&#x5730;&#x53EF;&#x4EE5;&#x4ECE; hESCs &#x751F;&#x6210; HPCs&#x3002;&#x6211;&#x4EEC;&#x7684;&#x7B56;&#x7565;&#x53EF;&#x4EE5;&#x63D0;&#x4F9B;&#x4F53;&#x5916;&#x6A21;&#x578B;&#xFF0C;&#x7814;&#x7A76;&#x65E9;&#x671F;&#x4EBA;&#x7C7B;&#x9020;&#x8840;&#x7EC6;&#x80DE;&#x53D1;&#x80B2;&#x3002;

TGF&beta; &#x6291;&#x5236;&#x4F5C;&#x7528;&#x589E;&#x5F3A;&#x4E86;&#x4ECE;&#x4EBA;&#x7C7B;&#x80DA;&#x80CE;&#x5E72;&#x7EC6;&#x80DE;&#x7684;&#x9020;&#x8840;&#x8840;&#x7BA1;&#x5185;&#x76AE;&#x7EC6;&#x80DE;&#x4F7F;&#x7528;&#x9010;&#x6B65;&#x6218;&#x7565;&#x9020;&#x8840;&#x7956;&#x7EC6;&#x80DE;&#x7684;&#x751F;&#x6210;&#x3002;

Embryonic hematopoiesis is a complex process. Elucidating the mechanism regulating hematopoietic differentiation from pluripotent stem cells would allow us to establish a strategy to efficiently generate hematopoietic cells. However, the mechanism governing the generation of hematopoietic progenitors from human embryonic stem cells (hESCs) remains unknown. Here, on the basis of the emergence of CD43(+) hematopoietic cells from hemogenic endothelial (HE) cells, we demonstrated that VEGF was essential and sufficient, and that bFGF was synergistic with VEGF to specify the HE cells and the subsequent transition into CD43(+) hematopoietic cells. Significantly, we identified TGFÎ² as a novel signal to regulate hematopoietic development, as the TGFÎ² inhibitor SB 431542 significantly promoted the transition from HE cells into CD43(+) hematopoietic progenitor cells (HPCs) during hESC differentiation. By defining these critical signaling factors during hematopoietic differentiation, we can efficiently generate HPCs from hESCs. Our strategy could offer an in vitro model to study early human hematopoietic development.

TGFÎ² inhibition enhances the generation of hematopoietic progenitors from human ES cell-derived hemogenic endothelial cells using a stepwise strategy.

Embryonnaire h&#xE9;matopo&#xEF;&#xE8;se est un processus complexe. &#xC9;lucider le m&#xE9;canisme de r&#xE9;gulation de la diff&#xE9;renciation h&#xE9;matopo&#xEF;&#xE9;tique des cellules souches pluripotentes nous permettrait d&#x26;#39;&#xE9;tablir une strat&#xE9;gie pour g&#xE9;n&#xE9;rer efficacement des cellules h&#xE9;matopo&#xEF;&#xE9;tiques. Cependant, le m&#xE9;canisme r&#xE9;gissant la production de prog&#xE9;niteurs h&#xE9;matopo&#xEF;&#xE9;tiques &#xE0; partir de cellules souches embryonnaires humaines (CSEh) reste inconnue. Ici, sur la base de l&#x26;#39;&#xE9;mergence de CD43 (+) des cellules h&#xE9;matopo&#xEF;&#xE9;tiques de l&#x26;#39;endoth&#xE9;lium hemogenic (SE) des cellules, nous avons d&#xE9;montr&#xE9; que le VEGF est essentiel et suffisant, et que le bFGF a &#xE9;t&#xE9; synergique avec le VEGF pour sp&#xE9;cifier les cellules ES et la transition subs&#xE9;quente en CD43 (+) des cellules h&#xE9;matopo&#xEF;&#xE9;tiques. De mani&#xE8;re significative, nous avons identifi&#xE9; TGF&#xDF; comme un signal roman pour r&#xE9;glementer le d&#xE9;veloppement h&#xE9;matopo&#xEF;&#xE9;tique, comme l&#x26;#39;inhibiteur de TGF SB 431542 consid&#xE9;rablement favoris&#xE9; le passage de l&#x26;#39;HE cellules en CD43 (+) des cellules prog&#xE9;nitrices h&#xE9;matopo&#xEF;&#xE9;tiques (HPC) au cours de la diff&#xE9;renciation des CSEh. En d&#xE9;finissant ces facteurs critiques de signalisation lors de la diff&#xE9;renciation h&#xE9;matopo&#xEF;&#xE9;tique, nous pouvons g&#xE9;n&#xE9;rer efficacement des CPS de CSEh. Notre strat&#xE9;gie pourrait offrir un mod&#xE8;le in vitro pour &#xE9;tudier le d&#xE9;veloppement pr&#xE9;coce h&#xE9;matopo&#xEF;&#xE9;tiques humaines.

Inhibition TGF&#xDF; Am&#xE9;liore la g&#xE9;n&#xE9;ration de prog&#xE9;niteurs h&#xE9;matopo&#xEF;&#xE9;tiques de ES humaines d&#xE9;riv&#xE9;es de cellules Hemogenic cellules endoth&#xE9;liales l&#x26;#39;aide d&#x26;#39;une strat&#xE9;gie par &#xE9;tapes

Embryonale Blutbildung ist ein komplexer Prozess. Elucidating des Mechanismus zur Regelung von h&auml;matopoetischen Differenzierung von pluripotenten Stammzellen w&uuml;rde uns eine Strategie um effizient generieren blutbildender Zellen erm&ouml;glichen. Der Mechanismus f&uuml;r die Generierung von h&auml;matopoetischen Vorl&auml;uferzellen aus humanen embryonalen Stammzellen (Zellkulturen) bleibt jedoch unbekannt. Hier auf der Grundlage der Entstehung CD43(+) blutbildender Zellen aus Hemogenic endothelial Zellen (HE), wir bewiesen, dass VEGF notwendig und ausreichend war, und diese bFGF synergistische mit VEGF die HE-Zellen und den sp&auml;teren &Uuml;bergang in CD43(+) blutbildender Zellen angeben. Bezeichnenderweise haben wir TGF&beta;, wie ein Roman Signal, h&auml;matopoetischen Entwicklung, zu regulieren, wie die TGF&beta;-Inhibitor SB 431542 deutlich den &Uuml;bergang vom er gef&ouml;rdert w&auml;hrend hESC Differenzierung in CD43(+) h&auml;matopoetischen Vorl&auml;uferzellen (HPCs) Zellen identifiziert. Definieren Sie diese kritischen signalisierende Faktoren w&auml;hrend der h&auml;matopoetischen Differenzierung, k&ouml;nnen wir effizient HPCs aus Zellkulturen generieren. Unsere Strategie k&ouml;nnte eine in-vitro-Modell, um fr&uuml;he menschliche Entwicklung h&auml;matopoetischen studieren anbieten.

TGF&beta; Hemmung erh&ouml;ht die Generation von h&auml;matopoetischen Vorl&auml;uferzellen aus menschlichen ES Cell-derived Hemogenic endothelial Zellen mit einer schrittweise-Strategie.

Emopoiesi embrionale sono un processo complesso. Chiarire il meccanismo di regolazione della differenziazione ematopoietiche da cellule staminali pluripotenti ci permetterebbe di stabilire una strategia per generare in modo efficiente le cellule ematopoietiche. Tuttavia, il meccanismo che regolano la generazione dei progenitori ematopoietici da cellule staminali embrionali umane (hESC) rimane sconosciuto. Qui, sulla base di emersione delle cellule ematopoietiche CD43(+) da hemogenic (egli) cellule endoteliali, abbiamo dimostrato che VEGF &egrave; stato essenziale e sufficiente, e che bFGF era sinergica con VEGF per specificare le celle di lui e il passaggio successivo in cellule ematopoietiche CD43(+). Significativamente, abbiamo identificato TGF&beta; come un segnale di romanzo a regolare lo sviluppo ematopoietico, come l&#39;inibitore TGF&beta; SB 431542 promosso significativamente la transizione da lui le cellule in CD43(+) le cellule progenitrici ematopoietiche (HPC) durante la differenziazione hESC. Con la definizione di questi fattori critici di segnalazione durante la differenziazione ematopoietiche, possiamo generare HPCs efficientemente da hESC. La nostra strategia &egrave; in grado di offrire un modello in vitro per lo studio precoce sviluppo ematopoietico umano.

Inibizione di TGF&beta; migliora la generazione dei progenitori ematopoietici da ES hemogenic derivato da cellule endoteliali cellule umane usando una strategia graduale.

&#x80CE;&#x5150;&#x306E;&#x9020;&#x8840;&#x306F;&#x8907;&#x96D1;&#x306A;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x3067;&#x3059;&#x3002;&#x591A;&#x80FD;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x9020;&#x8840;&#x7CFB;&#x3078;&#x306E;&#x5206;&#x5316;&#x3092;&#x8ABF;&#x7BC0;&#x3059;&#x308B;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x3092;&#x89E3;&#x660E;&#x3059;&#x308B;&#x3053;&#x3068;&#x306F;&#x79C1;&#x305F;&#x3061;&#x306F;&#x52B9;&#x7387;&#x7684;&#x306A;&#x9020;&#x8840;&#x7D30;&#x80DE;&#x3092;&#x751F;&#x6210;&#x3059;&#x308B;&#x305F;&#x3081;&#x306E;&#x6226;&#x7565;&#x3092;&#x78BA;&#x7ACB;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x308B;&#x3067;&#x3057;&#x3087;&#x3046;&#x3002;&#x3057;&#x304B;&#x3057;&#x3001;&#x30D2;&#x30C8;&#x80DA;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#xFF08;&#x30D2;&#x30C8;ES&#x7D30;&#x80DE;&#xFF09;&#x304B;&#x3089;&#x9020;&#x8840;&#x524D;&#x99C6;&#x7D30;&#x80DE;&#x306E;&#x751F;&#x6210;&#x3092;&#x652F;&#x914D;&#x3059;&#x308B;&#x30E1;&#x30AB;&#x30CB;&#x30BA;&#x30E0;&#x306F;&#x4E0D;&#x660E;&#x3067;&#x3042;&#x308B;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;hemogenic&#xFF08;HE&#xFF09;&#x5185;&#x76AE;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x306E;CD43&#xFF08;+&#xFF09;&#x9020;&#x8840;&#x7D30;&#x80DE;&#x306E;&#x51FA;&#x73FE;&#x306B;&#x57FA;&#x3065;&#x3044;&#x3066;&#x3001;&#x6211;&#x3005;&#x306F;VEGF&#x304C;&#x4E0D;&#x53EF;&#x6B20;&#x3068;&#x5341;&#x5206;&#x3067;&#x3042;&#x3063;&#x305F;&#x3001;&#x305D;&#x306E;bFGF&#x304C;HE&#x7D30;&#x80DE;&#x3068;CD43&#x3078;&#x306E;&#x305D;&#x306E;&#x5F8C;&#x306E;&#x5909;&#x9077;&#x3092;&#x6307;&#x5B9A;&#x3059;&#x308B;&#x306B;&#x306F;&#x3001;VEGF&#x3068;&#x306E;&#x76F8;&#x4E57;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x305F;&#xFF08;+&#xFF09;&#x9020;&#x8840;&#x7D30;&#x80DE;&#x3002;&#x8457;&#x3057;&#x304F;&#x3001;&#x6211;&#x3005;&#x306F;TGF-&#x3B2;&#x963B;&#x5BB3;&#x5264;&#x3068;&#x3057;&#x3066;SB 431542&#x306F;&#x5927;&#x5E45;&#x306B;HE&#x306F;&#x3001;&#x7D30;&#x80DE;&#x304B;&#x3089;CD43&#xFF08;+&#xFF09;hESC&#x306E;&#x5206;&#x5316;&#x306B;&#x304A;&#x3051;&#x308B;&#x9020;&#x8840;&#x524D;&#x99C6;&#x7D30;&#x80DE;&#xFF08;HPCS&#xFF09;&#x3078;&#x306E;&#x79FB;&#x884C;&#x3092;&#x4FC3;&#x9032;&#x3057;&#x3001;&#x9020;&#x8840;&#x767A;&#x751F;&#x3092;&#x5236;&#x5FA1;&#x3059;&#x308B;&#x305F;&#x3081;&#x306E;&#x65B0;&#x3057;&#x3044;&#x4FE1;&#x53F7;&#x3068;&#x3057;&#x3066;TGF&#x3B2;&#x3092;&#x540C;&#x5B9A;&#x3057;&#x305F;&#x3002;&#x9020;&#x8840;&#x5206;&#x5316;&#x306E;&#x904E;&#x7A0B;&#x3067;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x91CD;&#x8981;&#x306A;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x56E0;&#x5B50;&#x3092;&#x5B9A;&#x7FA9;&#x3059;&#x308B;&#x3053;&#x3068;&#x306B;&#x3088;&#x3063;&#x3066;&#x3001;&#x6211;&#x3005;&#x306F;&#x52B9;&#x7387;&#x7684;&#x306B;&#x30D2;&#x30C8;ES&#x7D30;&#x80DE;&#x304B;&#x3089;HPC&#x306E;&#x3092;&#x751F;&#x6210;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x5F53;&#x793E;&#x306E;&#x6226;&#x7565;&#x306F;&#x3001;&#x521D;&#x671F;&#x306E;&#x30D2;&#x30C8;&#x9020;&#x8840;&#x958B;&#x767A;&#x3092;&#x7814;&#x7A76;&#x3059;&#x308B;&#x305F;&#x3081;&#x306E;in vitro&#x30E2;&#x30C7;&#x30EB;&#x3067;&#x63D0;&#x4F9B;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;

TGF&#x3B2;&#x963B;&#x5BB3;&#x306F;&#x3001;&#x6BB5;&#x968E;&#x7684;&#x6226;&#x7565;&#x3092;&#x7528;&#x3044;&#x305F;&#x30D2;&#x30C8;ES&#x7D30;&#x80DE;&#x7531;&#x6765;&#x306E;Hemogenic&#x5185;&#x76AE;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x306E;&#x9020;&#x8840;&#x524D;&#x99C6;&#x7D30;&#x80DE;&#x306E;&#x751F;&#x6210;&#x3092;&#x5F37;&#x5316;

&#xBC30;&#xC544; &#xC870; &#xBCF5;&#xC7A1; &#xD55C; &#xACFC;&#xC815; &#xC774;&#xB2E4;&#xC785;&#xB2C8;&#xB2E4;. Pluripotent &#xC904;&#xAE30; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xC870; &#xD608; &#xBD84;&#xD654; &#xC870;&#xC808; &#xBA54;&#xCEE4;&#xB2C8;&#xC998; elucidating &#xC6B0;&#xB9AC;&#xAC00; &#xD6A8;&#xC728;&#xC801;&#xC73C;&#xB85C; &#xC870; &#xD608; &#xC138;&#xD3EC;&#xB97C; &#xC0DD;&#xC131; &#xD558;&#xB294; &#xC804;&#xB7B5;&#xC744; &#xC218;&#xB9BD; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;, &#xC778;&#xAC04; &#xBC30;&#xC544; &#xC904;&#xAE30; &#xC138;&#xD3EC; (hESCs)&#xC5D0;&#xC11C; &#xC870; &#xD608; progenitors&#xC758; &#xC0DD;&#xC131;&#xC744; &#xC81C;&#xC5B4; &#xD558;&#xB294; &#xBA54;&#xCEE4;&#xB2C8;&#xC998; &#xC54C; &#xC218; &#xC5C6;&#xB294; &#xB0A8;&#xC544; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30;, hemogenic&#xC5D0;&#xC11C; CD43(+) &#xC870; &#xD608; &#xC138;&#xD3EC;&#xC758; &#xCD9C;&#xD604; &#xADFC;&#xAC70; (&#xADF8;&#xAC00;) &#xB0B4; &#xD53C; &#xC138;&#xD3EC;, VEGF&#xB294; &#xC911;&#xC694; &#xD558; &#xACE0; &#xCDA9;&#xBD84; &#xD55C;, &#xADF8;&#xB9AC;&#xACE0; &#xADF8; bFGF &#xADF8; &#xC140;&#xACFC; CD43(+) &#xC870; &#xD608; &#xC138;&#xD3EC;&#xB85C; &#xD6C4;&#xC18D; &#xC804;&#xD658;&#xC744; &#xC9C0;&#xC815; &#xD558;&#xB294; VEGF&#xC640; &#xC2DC;&#xB108;&#xC9C0;&#xB294; &#xC6B0;&#xB9AC; &#xC2DC;&#xC5F0;. HESC &#xCC28;&#xBCC4;&#xD654; &#xD558;&#xB294; &#xB3D9;&#xC548; CD43(+) &#xC870;&#xC0C1; &#xC870; &#xD608; &#xC138;&#xD3EC; (HPCs)&#xB85C; &#xC138;&#xD3EC;&#xAC00; &#xC18C;&#xC124; &#xC2E0;&#xD638; TGF&beta; &#xC5B5;&#xC81C;&#xC81C; SB 431542 &#xD06C;&#xAC8C; &#xC2B9;&#xC9C4;&#xC5D0;&#xC11C; &#xADF8;&#xB294; &#xC804;&#xD658; hematopoietic &#xAC1C;&#xBC1C; &#xADDC;&#xC81C;&#xB97C; &#xD06C;&#xAC8C;, &#xC6B0;&#xB9AC; TGF&beta; &#xC2DD;&#xBCC4;. Hematopoietic &#xCC28;&#xBCC4;&#xD654; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xC774;&#xB7EC;&#xD55C; &#xC911;&#xC694; &#xD55C; &#xC2E0;&#xD638; &#xC694;&#xC18C;&#xB97C; &#xC815;&#xC758; &#xD558; &#xC5EC; &#xC6B0;&#xB9AC; Hescs&#xC5D0;&#xC11C; Hpcs&#xB97C; &#xD6A8;&#xC728;&#xC801;&#xC73C;&#xB85C; &#xC0DD;&#xC131;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC;&#xC758; &#xC804;&#xB7B5;&#xC740; &#xCD08;&#xAE30; &#xC778;&#xAC04; &#xC870; &#xD608; &#xAC1C;&#xBC1C;&#xC744; &#xACF5;&#xBD80; &#xD558;&#xB294; &#xCCB4; &#xC678; &#xBAA8;&#xB378;&#xC744; &#xC81C;&#xACF5;&#xD560; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;.

TGF&beta; &#xC5B5;&#xC81C; &#xC778;&#xAC04;&#xC758; ES &#xC138;&#xD3EC; &#xC720;&#xB798; hemogenic &#xB0B4; &#xD53C; &#xC138;&#xD3EC; stepwise &#xC804;&#xB7B5;&#xC744; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xC870; &#xD608; progenitors&#xC758; &#xC0DD;&#xC131;&#xC744; &#xD5A5;&#xC0C1; &#xC2DC;&#xD0B5;&#xB2C8;&#xB2E4;.

Embrionario hematopoyesis es un proceso complejo. Dilucidar el mecanismo de regulaci&#xF3;n de la diferenciaci&#xF3;n hematopoy&#xE9;tica de las c&#xE9;lulas madre pluripotentes nos permitir&#xED;a establecer una estrategia para generar eficientemente las c&#xE9;lulas hematopoy&#xE9;ticas. Sin embargo, el mecanismo que rige la generaci&#xF3;n de progenitores hematopoy&#xE9;ticos de c&#xE9;lulas madre embrionarias humanas (hESCs) sigue siendo desconocido. Aqu&#xED;, sobre la base de la aparici&#xF3;n de CD43 (+) de las c&#xE9;lulas hematopoy&#xE9;ticas endotelial hemogenic (HE) c&#xE9;lulas, hemos demostrado que el VEGF es esencial y suficiente, y que el bFGF es sin&#xE9;rgica con el VEGF para especificar las c&#xE9;lulas HE y la transici&#xF3;n posterior en CD43 (+) las c&#xE9;lulas hematopoy&#xE9;ticas. Es significativo que hemos identificado como una se&#xF1;al de TGF novela para regular el desarrollo hematopoy&#xE9;tico, como el inhibidor de TGF SB 431542 promovido considerablemente la transici&#xF3;n de HE c&#xE9;lulas CD43 en la (+) las c&#xE9;lulas progenitoras hematopoy&#xE9;ticas (HPC) durante la diferenciaci&#xF3;n hESC. Mediante la definici&#xF3;n de estos factores cr&#xED;ticos de se&#xF1;alizaci&#xF3;n durante la diferenciaci&#xF3;n hematopoy&#xE9;tica, que de manera eficiente puede generar HPC de hESCs. Nuestra estrategia podr&#xED;a ofrecer un modelo in vitro para estudiar el desarrollo temprano hematopoy&#xE9;tico humano.

La inhibici&#xF3;n TGF&#xDF; realza la generaci&#xF3;n de progenitores hematopoy&#xE9;ticos de la c&#xE9;lula madre embrionarias humanas derivadas de las c&#xE9;lulas endoteliales Hemogenic Utilizando una estrategia paso a paso

We have demonstrated that c-Src suppression inhibited the epithelial to mesenchymal transition in human breast cancer cells. Here, we investigated the role of c-Src on the cell cycle progression using siRNAs and small molecule inhibitor 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2). Western blot analysis demonstrated the down-regulation of cyclin D1 and cyclin E and up-regulation of p27 Kip1 after c-Src suppression by PP2. Incubation of cells in the presence of PP2 significantly blocked the phosphorylation of extracellular signal-regulated kinases 1/2 (ERK1/2), protein kinase B (AKT), and glycogen synthase kinase 3 beta (GSK3β). Specific pharmacological inhibitors of MEK1/2/ERK1/2 and phosphatidylinositide 3-kinase/AKT pathways were used to demonstrate the relationship between the signal cascade and cell cycle proteins expression. The expression of cyclin D1 and cyclin E were decreased after inhibition of ERK1/2 or AKT activity, whereas the p27 Kip1 expression was increased. In addition, knockdown of c-Src by siRNAs reduced cell proliferation and phosphorylation of ERK1/2, AKT, and GSK3β. After c-Src depletion by siRNAs, we observed significant down-regulation of cyclin D1 and cyclin E, and up-regulation of p27 Kip1. These results suggest that c-Src suppression by PP2 or siRNAs may regulate the progression of cell cycle through AKT/GSK3β and ERK1/2 pathways. 

c-Src regulates cell cycle proteins expression through protein kinase B/glycogen synthase kinase 3 beta and extracellular signal-regulated kinases 1/2 pathways in MCF-7 cells.

The cells with mammary repopulating capability can achieve mammary gland morphogenesis in a suitable cellular microenvironment. Using cell surface markers of CD24, CD29 and CD49f, mouse mammary repopulating unit (MRU) has been identified in adult mammary epithelium and late embryonic mammary bud epithelium. However, embryonic MRU remains to be fully characterized at earlier mammary anlagen stage. Here we isolated discrete populations of E14.5 mouse mammary anlagen cells. Only Lin(-)CD24(med)CD29(+) cell population was predicted as E14.5 MRU by examining their capacities of forming mammosphere and repopulating cleared mammary fat pad in vivo. However, when we characterized gene expressions of this E14.5 cell population by comparing with adult mouse MRU (Lin(-)CD24(+)CD29(hi)), the gene profiling of these two cell populations exhibited great differences. Real-time PCR and immunostaining assays uncovered that E14.5 Lin(-)CD24(med)CD29(+) cell population was a heterogeneous stroma-enriched cell population. Then, limiting dilutions and single-cell assays also confirmed that E14.5 Lin(-)CD24(med)CD29(+) cell population possessed low proportion of stem cells. In summary, heterogeneous Lin(-)CD24(med)CD29(+) cell population exhibited mammary repopulating ability in E14.5 mammary anlagen, implying that only suitable mammary stroma could enable mammary gland morphogenesis, which relied on the interaction between rare stem cells and microenvironment. 

Liying Du

School of Life Sciences

Peking University

Nucleofection and In Vivo Propagation of Chicken Eimeria Parasites