modeling-hepatitis-b-virus-infection-in-non-hepatic-293t-ne-3nrs-cells

Modeling Hepatitis B Virus Infection in Non-Hepatic 293T-NE-3NRs Cells

HBV mainly infects human hepatocytes, but it has also been found to infect extrahepatic tissues such as kidney and testis. Nonetheless, cell-based HBV ...

an-efficient-method-for-directed-hepatocyte-like-cell-induction-from

An Efficient Method for Directed Hepatocyte-Like Cell Induction from Human Embryonic Stem Cells

The potential functions of hepatocyte-like cells (HLCs) derived from human embryonic stem cells (hESCs) hold great promise for disease modeling and drug ...

Shantou University Medical College

&#x4E59;&#x578B;&#x809D;&#x708E;&#x75C5;&#x6BD2; (HBV)&#x3001; &#x4E19;&#x578B;&#x809D;&#x708E;&#x75C5;&#x6BD2; (HCV) &#x611F;&#x67D3;&#x7684;&#x7ED3;&#x679C;&#x5F02;&#x6784;&#x4ECE;&#x6025;&#x6027;&#x65E0;&#x75C7;&#x72B6;&#x611F;&#x67D3;&#x5BFC;&#x81F4;&#x809D;&#x786C;&#x5316;&#x548C;&#x809D;&#x7EC6;&#x80DE;&#x764C; (HCC) &#x7684;&#x6162;&#x6027;&#x611F;&#x67D3;&#x3002;&#x4F7F;&#x7528;&#x52A8;&#x7269;&#x809D;&#x7EC6;&#x80DE;&#x5728;&#x4F53;&#x5916;&#x6A21;&#x578B;&#xFF0C;&#x4EBA;&#x7C7B;&#x809D;&#x764C;&#x7EC6;&#x80DE;&#x682A;&#x6216;&#x4F53;&#x5185;&#x8F6C;&#x57FA;&#x56E0;&#x5C0F;&#x9F20;&#x6A21;&#x578B;&#x6709;&#x52A9;&#x4E8E;&#x4E3A;&#x6570;&#x4F17;&#x591A;&#x4E86;&#x89E3;&#x4E59;&#x578B;&#x809D;&#x708E;&#x75C5;&#x6BD2;&#x548C;&#x4E19;&#x578B;&#x809D;&#x708E;&#x7684;&#x53D1;&#x75C5;&#x673A;&#x5236;&#x3002;&#x6240;&#x4F5C;&#x7684;&#x91CD;&#x7EC4;&#x4EBA;&#x539F;&#x53D1;&#x6027;&#x809D;&#x7EC6;&#x80DE;&#x7684;&#x514D;&#x75AB;&#x7F3A;&#x9677;&#x7684;&#x5C0F;&#x9F20;&#x809D;&#x810F;&#x4E2D;&#x7684;&#x4EBA;&#x6027;&#x5316;&#x7684;&#x9F20;&#x6807;&#x6A21;&#x578B;&#x63D0;&#x4F9B;&#x4E86;&#x65B0;&#x9896;&#x7684;&#x5B9E;&#x9A8C;&#x673A;&#x4F1A;&#xFF0C;&#x6A21;&#x4EFF;&#x4EBA;&#x7C7B;&#x809D;&#x7EC6;&#x80DE;&#x5728;&#x4F53;&#x5185;&#x751F;&#x957F;&#x3002;&#x4EBA;&#x539F;&#x53D1;&#x6027;&#x809D;&#x7EC6;&#x80DE;&#x83B7;&#x5F97;&#x6709;&#x9650;&#x4E0D;&#x5F97;&#x4E0D;&#x5BFB;&#x627E;&#x5176;&#x4ED6;&#x7EC6;&#x80DE;&#x7684;&#x6765;&#x6E90;&#xFF0C;&#x5982;&#x591A;&#x80FD;&#x5E72;&#x7EC6;&#x80DE;&#x3002;&#x4EBA;&#x7C7B;&#x80DA;&#x80CE;&#x5E72;&#x7EC6;&#x80DE; (hESCs) &#x5177;&#x6709;&#x81EA;&#x6211;&#x66F4;&#x65B0;&#x548C;&#x591A;&#x6F5C;&#x80FD;&#x7684;&#x529F;&#x80FD;&#x548C;&#x6240;&#x6709;&#x4E09;&#x4E2A;&#x80DA;&#x5C42;&#xFF0C;&#x5305;&#x62EC;&#x809D;&#x7EC6;&#x80DE;&#x7684;&#x5206;&#x5316;&#x3002;Humaninduced &#x591A;&#x80FD;&#x5E72;&#x7EC6;&#x80DE; (&#x9B54;&#x9B3C;) &#x6765;&#x81EA;&#x75C5;&#x4EBA;&#x7684;&#x6216;&#x4E2A;&#x4EBA;&#x7684;&#x7EC6;&#x80DE;&#x63D0;&#x4F9B;&#x4E86;&#x65B0;&#x7684;&#x673A;&#x4F1A;&#xFF0C;&#x4EE5;&#x751F;&#x6210;&#x809D;&#x7EC6;&#x80DE;&#x6837;&#x7EC6;&#x80DE;&#x4E0E;&#x754C;&#x5B9A;&#x7684;&#x9057;&#x4F20;&#x7EC4;&#x6210;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#xFF0C;&#x6211;&#x4EEC;&#x4F1A;&#x68C0;&#x8BA8;&#x5F53;&#x524D;&#x89C6;&#x89D2;&#x7684;&#x6A21;&#x578B;&#x7528;&#x4E8E;&#x4E59;&#x578B;&#x809D;&#x708E;&#x75C5;&#x6BD2;&#x548C;&#x4E19;&#x578B;&#x809D;&#x708E;&#x75C5;&#x6BD2;&#x7684;&#x7814;&#x7A76;&#xFF0C;&#x5E76;&#x4ECB;&#x7ECD;&#x4F7F;&#x7528;&#x4EBA;&#x7C7B;&#x9B54;&#x9B3C;&#x7684;&#x4E2A;&#x6027;&#x5316;&#x7684;&#x9F20;&#x6807;&#x6A21;&#x578B;&#x3002;&#x8FD9;&#x79CD;&#x65B0;&#x578B;&#x9F20;&#x6807;&#x6A21;&#x5F0F;&#x5C06;&#x6709;&#x52A9;&#x4E8E;&#x4E59;&#x578B;&#x809D;&#x708E;&#x75C5;&#x6BD2;&#x548C;&#x4E19;&#x578B;&#x809D;&#x708E;&#x75C5;&#x6BD2;&#x7684;&#x4EBA;&#x7C7B;&#x809D;&#x7EC6;&#x80DE;&#xFF0C;&#x4EE5;&#x53CA;&#x63A2;&#x8BA8;&#x5BF9;&#x809D;&#x7EC6;&#x80DE;&#x5BF9;&#x4E59;&#x578B;&#x809D;&#x708E;&#x75C5;&#x6BD2;&#x548C;&#x4E19;&#x578B;&#x809D;&#x708E;&#x75C5;&#x6BD2;&#x6613;&#x611F;&#x6027;&#x7684;&#x9057;&#x4F20;&#x5F71;&#x54CD;&#x7684;&#x76F4;&#x63A5;&#x8C03;&#x67E5;&#x3002;

&#x4EBA;&#x6027;&#x5316;&#x7684;&#x4E59;&#x578B;&#x809D;&#x708E;&#x75C5;&#x6BD2;&#x548C;&#x4E19;&#x578B;&#x809D;&#x708E;&#x75C5;&#x6BD2;&#x4F7F;&#x7528;&#x4EBA;&#x7C7B;&#x8BF1;&#x5BFC;&#x591A;&#x80FD;&#x5E72;&#x7EC6;&#x80DE;&#x7684;&#x5C0F;&#x9F20;&#x6A21;&#x578B;&#x3002;

Infection of hepatitis B virus (HBV) and hepatitis C virus (HCV) results in heterogeneous outcomes from acute asymptomatic infection to chronic infection leading to cirrhosis and hepatocellular carcinoma (HCC). In vitro models using animal hepatocytes, human HCC cell lines, or in vivo transgenic mouse models have contributed invaluably to understanding the pathogenesis of HBV and HCV. A humanized mouse model made by reconstitution of human primary hepatocytes in the liver of the immunodeficient mouse provides a novel experimental opportunity which mimics the in vivo growth of the human hepatocytes. The limited access to primary human hepatocytes necessitated the search for other cellular sources, such as pluripotent stem cells. Human embryonic stem cells (hESCs) have the features of self-renewal and pluripotency and differentiate into cells of all three germ layers, including hepatocytes. Humaninduced pluripotent stem cells (iPSCs) derived from the patient's or individual's own cells provide a novel opportunity to generate hepatocyte-like cells with the defined genetic composition. Here, we will review the current perspective of the models used for HBV and HCV study, and introduce the personalized mouse model using human iPSCs. This novel mouse model will facilitate the direct investigation of HBV and HCV in human hepatocytes as well as probing the genetic influence on the susceptibility of hepatocytes to HBV and HCV.

Humanized murine model for HBV and HCV using human induced pluripotent stem cells.

L&#x26;#39;infection de l&#x26;#39;h&#xE9;patite B (VHB) et l&#x26;#39;h&#xE9;patite C (VHC) aboutit &#xE0; des r&#xE9;sultats h&#xE9;t&#xE9;rog&#xE8;nes de l&#x26;#39;infection asymptomatique &#xE0; l&#x26;#39;infection aigu&#xEB; chronique conduisant &#xE0; la cirrhose et le carcinome h&#xE9;patocellulaire (CHC). Dans les mod&#xE8;les in vitro utilisant des h&#xE9;patocytes des animaux, lign&#xE9;es cellulaires humaines de carcinome h&#xE9;patocellulaire, ou dans des mod&#xE8;les in vivo de souris transg&#xE9;niques ont contribu&#xE9; de fa&#xE7;on inestimable &#xE0; la compr&#xE9;hension de la pathogen&#xE8;se de VHB et le VHC. Un mod&#xE8;le de souris humanis&#xE9;e par la reconstitution des h&#xE9;patocytes primaires humains dans le foie de la souris immunod&#xE9;ficientes offre une occasion roman exp&#xE9;rimental qui imite la croissance in vivo des h&#xE9;patocytes humains. L&#x26;#39;acc&#xE8;s limit&#xE9; aux h&#xE9;patocytes humains primaires a n&#xE9;cessit&#xE9; la recherche d&#x26;#39;autres sources cellulaires, telles que les cellules souches pluripotentes. Cellules souches embryonnaires humaines (CSEh) pr&#xE9;sentent les caract&#xE9;ristiques de l&#x26;#39;auto-renouvellement et la pluripotence et se diff&#xE9;rencier en cellules de tous les trois feuillets embryonnaires, y compris les h&#xE9;patocytes. Anthropog&#xE9;niques cellules souches pluripotentes (CISP) provenant de du patient ou de ses propres cellules individuelles de fournir une nouvelle opportunit&#xE9; de g&#xE9;n&#xE9;rer des h&#xE9;patocytes comme les cellules avec la composition d&#xE9;finie g&#xE9;n&#xE9;tique. Ici, nous allons examiner la perspective actuelle des mod&#xE8;les utilis&#xE9;s pour le VHB et le VHC &#xE9;tude, et d&#x26;#39;introduire le mod&#xE8;le de souris personnalis&#xE9; &#xE0; l&#x26;#39;aide CSPi humaines. Ce nouveau mod&#xE8;le de souris sera de faciliter l&#x26;#39;enqu&#xEA;te directe du VHB et du VHC dans les h&#xE9;patocytes humains, ainsi que sonder l&#x26;#39;influence g&#xE9;n&#xE9;tique sur la sensibilit&#xE9; des h&#xE9;patocytes au VHB et le VHC.

Mod&#xE8;le murin humanis&#xE9; pour le VHB et VHC &#xE0; l&#x26;#39;aide de l&#x26;#39;homme cellules souches pluripotentes induites

Infektion der Hepatitis B-Virus (HBV) und Hepatitis-C-Virus (HCV) f&uuml;hrt heterogene Ergebnisse durch akute asymptomatische Infektion f&uuml;r chronische Infektion f&uuml;hrt zu Leberzirrhose und Hepatozellul&auml;res Karzinom (HCC). In-vitro-Modelle mit tierische Hepatozyten, trugen menschliche HCC-Zelllinien oder in-vivo transgener Mausmodelle unsch&auml;tzbar zum besseren Verst&auml;ndnis der Pathogenese der HBV und HCV. Einem humanisierten Mausmodell gemacht durch R&uuml;ckverd&uuml;nnung von menschlichen prim&auml;re Hepatozyten in der Leber der immungeschw&auml;chte Maus bietet eine neuartige experimentelle Gelegenheit die in-vivo Wachstum der menschlichen Hepatozyten imitiert. Der begrenzte Zugang zu prim&auml;ren menschlichen Hepatozyten erforderten die Suche nach anderen zellularen Quellen wie pluripotenten Stammzellen. Menschlicher embryonaler Stammzellen (Zellkulturen) haben die Merkmale der Selbsterneuerung und Pluripotenz und differenzieren sich in Zellen aller drei Keim-Schichten, einschlie&szlig;lich der Hepatozyten. Humaninduced pluripotenten Stammzellen (iPSCs) abgeleitet von der Patientin oder des einzelnen Zellen sorgen f&uuml;r eine neuartige M&ouml;glichkeit Hepatozyten-wie Zellen mit der definierten genetische Zusammensetzung. Hier werden wir die aktuelle Perspektive der Modelle f&uuml;r HBV und HCV verwendet zu studieren und personalisierte Mausmodell mit menschlichen iPSCs einzuf&uuml;hren. Diese neuartige Maus-Modell erleichtert die direkte Untersuchung von HBV und HCV in menschlichen Hepatozyten sowie die Sondierung des genetischen Einfluss auf die Empf&auml;nglichkeit von Hepatozyten, HBV und HCV.

Humanisierte murinen Modell f&uuml;r HBV und HCV menschlichen induzierten pluripotenten Stammzellen.

Infezione del virus dell&#39;epatite B (HBV) e virus dell&#39;epatite C (HCV) si traduce in risultati eterogenei da infezione asintomatica acuta di infezione cronica che porta a cirrosi e hepatocellular carcinoma (HCC). Modelli in vitro usando epatociti animali, linee di cellule HCC umani, o modelli murini transgenici in vivo hanno contribuito inestimabilmente alla comprensione della patogenesi dell&#39;HBV e HCV. Un modello murino umanizzato effettuato mediante ricostituzione di epatociti primari umani nel fegato del mouse immunodeficiente fornisce un&#39;occasione romanzo sperimentale che imita la crescita in vivo degli epatociti umani. L&#39;accesso limitato ai primari epatociti umani resa necessaria la ricerca di altre fonti cellulari, come le cellule staminali pluripotenti. Cellule staminali embrionali umane (hESC) hanno le caratteristiche di auto-rinnovamento e pluripotenza e differenziarsi in cellule di tutti i tre strati di germe, compresi gli epatociti. Humaninduced pluripotent stem cells (iPSCs) derivato da del paziente o le cellule dell&#39;individuo forniscono una romanzo opportunit&agrave; di generare degli epatociti-come le cellule con la composizione genetica definita. Qui, noi esamineremo le attuali prospettive dei modelli usati per HBV e HCV e studiare e introdurre il modello del mouse personalizzato utilizzando iPSCs umane. Questo modello di mouse romanzo faciliter&agrave; l&#39;indagine diretta di HBV e HCV in epatociti umani nonch&eacute; sondando l&#39;influenza genetica sulla suscettibilit&agrave; di epatociti per HBV e HCV.

Modello murino umanizzato per HBV e HCV utilizzando cellule staminali umane pluripotenti indotte.

B&#x578B;&#x809D;&#x708E;&#x30A6;&#x30A4;&#x30EB;&#x30B9;&#xFF08;HBV&#xFF09;&#x304A;&#x3088;&#x3073;&#x6025;&#x6027;&#x7121;&#x75C7;&#x5019;&#x6027;&#x611F;&#x67D3;&#x304B;&#x3089;&#x809D;&#x786C;&#x5909;&#x3084;&#x809D;&#x7D30;&#x80DE;&#x764C;&#xFF08;HCC&#xFF09;&#x306B;&#x3064;&#x306A;&#x304C;&#x308B;&#x6162;&#x6027;&#x611F;&#x67D3;&#x75C7;&#x306B;&#x7570;&#x7A2E;&#x306E;&#x7D50;&#x679C;&#x3067;C&#x578B;&#x809D;&#x708E;&#x30A6;&#x30A4;&#x30EB;&#x30B9;&#xFF08;HCV&#xFF09;&#x306E;&#x611F;&#x67D3;&#x306E;&#x7D50;&#x679C;&#x3002;&#x52D5;&#x7269;&#x306E;&#x809D;&#x7D30;&#x80DE;&#x3001;&#x30D2;&#x30C8;HCC&#x7D30;&#x80DE;&#x682A;&#x3001;&#x307E;&#x305F;&#x306F;in vivo&#x3067;&#x306E;&#x30C8;&#x30E9;&#x30F3;&#x30B9;&#x30B8;&#x30A7;&#x30CB;&#x30C3;&#x30AF;&#x30DE;&#x30A6;&#x30B9;&#x30E2;&#x30C7;&#x30EB;&#x306F;&#x3001;HBV&#x304A;&#x3088;&#x3073;HCV&#x306E;&#x75C5;&#x56E0;&#x3092;&#x7406;&#x89E3;&#x3059;&#x308B;&#x975E;&#x5E38;&#x306B;&#x9AD8;&#x4FA1;&#x306B;&#x8CA2;&#x732E;&#x3057;&#x305F;&#x3092;&#x7528;&#x3044;&#x305F;in vitro&#x30E2;&#x30C7;&#x30EB;&#x3067;&#x3042;&#x308B;&#x3002;&#x514D;&#x75AB;&#x4E0D;&#x5168;&#x30DE;&#x30A6;&#x30B9;&#x306E;&#x809D;&#x81D3;&#x306B;&#x304A;&#x3051;&#x308B;&#x30D2;&#x30C8;&#x521D;&#x4EE3;&#x809D;&#x7D30;&#x80DE;&#x306E;&#x518D;&#x69CB;&#x6210;&#x306B;&#x3088;&#x3063;&#x3066;&#x884C;&#x308F;&#x308C;&#x305F;&#x30D2;&#x30C8;&#x5316;&#x30DE;&#x30A6;&#x30B9;&#x30E2;&#x30C7;&#x30EB;&#x306F;&#x30D2;&#x30C8;&#x809D;&#x7D30;&#x80DE;&#x306E;in vivo&#x3067;&#x306E;&#x6210;&#x9577;&#x3092;&#x6A21;&#x5023;&#x3057;&#x305F;&#x65B0;&#x305F;&#x306A;&#x5B9F;&#x9A13;&#x306E;&#x6A5F;&#x4F1A;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x30D7;&#x30E9;&#x30A4;&#x30DE;&#x30EA;&#x30FC;&#x30D2;&#x30C8;&#x809D;&#x7D30;&#x80DE;&#x3078;&#x306E;&#x30A2;&#x30AF;&#x30BB;&#x30B9;&#x304C;&#x5236;&#x9650;&#x3055;&#x308C;&#x3001;&#x3053;&#x306E;&#x3088;&#x3046;&#x306A;&#x591A;&#x80FD;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#x306E;&#x3088;&#x3046;&#x306A;&#x4ED6;&#x306E;&#x7D30;&#x80DE;&#x30BD;&#x30FC;&#x30B9;&#x306E;&#x63A2;&#x7D22;&#x3092;&#x5FC5;&#x8981;&#x3068;&#x3057;&#x305F;&#x3002;&#x30D2;&#x30C8;&#x80DA;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#xFF08;&#x30D2;&#x30C8;ES&#x7D30;&#x80DE;&#xFF09;&#x81EA;&#x5DF1;&#x518D;&#x751F;&#x304A;&#x3088;&#x3073;&#x591A;&#x80FD;&#x6027;&#x306E;&#x6A5F;&#x80FD;&#x3092;&#x6301;&#x3061;&#x3001;&#x809D;&#x7D30;&#x80DE;&#x3092;&#x542B;&#x3080;&#x3059;&#x3079;&#x3066;&#x306E;3&#x3064;&#x306E;&#x80DA;&#x8449;&#x306E;&#x7D30;&#x80DE;&#x306B;&#x5206;&#x5316;&#x3059;&#x308B;&#x3002;&#x60A3;&#x8005;&#x3055;&#x3093;&#x3084;&#x500B;&#x4EBA;&#x306E;&#x81EA;&#x8EAB;&#x306E;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x6D3E;&#x751F;&#x3057;&#x305F;Humaninduced&#x591A;&#x80FD;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#xFF08;&#x6027;IPSC&#xFF09;&#x304C;&#x5B9A;&#x7FA9;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x907A;&#x4F1D;&#x7684;&#x7D44;&#x6210;&#x3092;&#x6709;&#x3059;&#x308B;&#x809D;&#x7D30;&#x80DE;&#x69D8;&#x7D30;&#x80DE;&#x3092;&#x751F;&#x6210;&#x3059;&#x308B;&#x305F;&#x3081;&#x306E;&#x65B0;&#x305F;&#x306A;&#x6A5F;&#x4F1A;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;HBV&#x304A;&#x3088;&#x3073;HCV&#x7814;&#x7A76;&#x306E;&#x305F;&#x3081;&#x306B;&#x4F7F;&#x7528;&#x3055;&#x308C;&#x308B;&#x30E2;&#x30C7;&#x30EB;&#x306E;&#x73FE;&#x5728;&#x306E;&#x8996;&#x70B9;&#x3092;&#x78BA;&#x8A8D;&#x3057;&#x3001;&#x4EBA;&#x9593;&#x6027;IPSC&#x3092;&#x4F7F;&#x7528;&#x3057;&#x3066;&#x30D1;&#x30FC;&#x30BD;&#x30CA;&#x30E9;&#x30A4;&#x30BA;&#x3055;&#x308C;&#x305F;&#x30DE;&#x30A6;&#x30B9;&#x30E2;&#x30C7;&#x30EB;&#x3092;&#x3054;&#x7D39;&#x4ECB;&#x3057;&#x307E;&#x3059;&#x3002;&#x3053;&#x306E;&#x5C0F;&#x8AAC;&#x306E;&#x30DE;&#x30A6;&#x30B9;&#x30E2;&#x30C7;&#x30EB;&#x3067;&#x306F;&#x3001;HBV&#x304A;&#x3088;&#x3073;HCV&#x306E;&#x809D;&#x7D30;&#x80DE;&#x306E;&#x611F;&#x53D7;&#x6027;&#x306B;&#x907A;&#x4F1D;&#x7684;&#x5F71;&#x97FF;&#x3092;&#x63A2;&#x308B;&#x3060;&#x3051;&#x3067;&#x306A;&#x304F;&#x3001;&#x30D2;&#x30C8;&#x809D;&#x7D30;&#x80DE;&#x306B;HBV&#x304A;&#x3088;&#x3073;HCV&#x306E;&#x76F4;&#x63A5;&#x306E;&#x8ABF;&#x67FB;&#x3092;&#x5BB9;&#x6613;&#x306B;&#x3057;&#x307E;&#x3059;&#x3002;

&#x30D2;&#x30C8;&#x4EBA;&#x5DE5;&#x591A;&#x80FD;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#x3092;&#x7528;&#x3044;&#x305F;HBV&#x304A;&#x3088;&#x3073;HCV&#x306E;&#x30D2;&#x30C8;&#x5316;&#x30DE;&#x30A6;&#x30B9;&#x30E2;&#x30C7;&#x30EB;

B &#xD615; &#xAC04;&#xC5FC; &#xBC14;&#xC774;&#xB7EC;&#xC2A4; (HBV) &#xBC0F; C &#xD615; &#xAC04;&#xC5FC; &#xBC14;&#xC774;&#xB7EC;&#xC2A4; (HCV) &#xAC10;&#xC5FC; &#xACB0;&#xACFC; &#xAC04; &#xACBD; &#xBCC0; &#xAC04;&#xC138;&#xD3EC; &#xC554; (HCC)&#xB85C; &#xC774;&#xC5B4;&#xC9C0;&#xB294; &#xB9CC;&#xC131; &#xAC10;&#xC5FC;&#xC744; &#xAE09;&#xC131; &#xBB34; &#xC99D;&#xC0C1; &#xAC10;&#xC5FC;&#xC5D0;&#xC11C; &#xC720;&#xD615;&#xC774; &#xB2E4;&#xB978; &#xACB0;&#xACFC;. &#xCCB4; &#xC678; &#xBAA8;&#xB378; &#xB3D9;&#xBB3C; hepatocytes&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC;, &#xC778;&#xAC04;&#xC758; HCC &#xC140; &#xB77C;&#xC778; &#xB610;&#xB294; in vivo &#xC720;&#xC804;&#xC790; &#xBCC0;&#xD615; &#xB9C8;&#xC6B0;&#xC2A4; &#xBAA8;&#xB378; invaluably&#xC5D0; &#xAE30;&#xC5EC;&#xD55C; HBV&#xC640; HCV&#xC758; pathogenesis&#xC758; &#xC774;&#xD574;. Humanized &#xB9C8;&#xC6B0;&#xC2A4; &#xBAA8;&#xB378; immunodeficient &#xB9C8;&#xC6B0;&#xC2A4;&#xC758; &#xAC04;&#xC5D0;&#xC11C; &#xC778;&#xAC04;&#xC758; &#xAE30;&#xBCF8; hepatocytes&#xC758; &#xC7AC;&#xAD6C;&#xC131; &#xD558; &#xC5EC; &#xB9CC;&#xB4E0; &#xBAA8;&#xBC29;&#xD55C; &#xC778;&#xAC04;&#xC758; hepatocytes&#xC758; in vivo &#xC131;&#xC7A5; &#xC18C;&#xC124; &#xC2E4;&#xD5D8; &#xAE30;&#xD68C;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xAE30;&#xBCF8; &#xC778;&#xAC04;&#xC758; hepatocytes&#xC5D0; &#xC81C;&#xD55C; &#xB41C; &#xC561;&#xC138;&#xC2A4;&#xAC00; &#xD544;&#xC694;&#xD55C; pluripotent &#xC904;&#xAE30; &#xC138;&#xD3EC; &#xB4F1; &#xC138;&#xD3EC; &#xB2E4;&#xB978; &#xC18C;&#xC2A4;&#xC5D0; &#xB300; &#xD55C; &#xAC80;&#xC0C9;. &#xC778;&#xAC04; &#xBC30;&#xC544; &#xC904;&#xAE30; &#xC138;&#xD3EC; (hESCs) &#xC790;&#xAE30; &#xAC31;&#xC2E0;&#xACFC; pluripotency&#xC758; &#xAE30;&#xB2A5;&#xC744; &#xAC16;&#xACE0; &#xC788;&#xC73C;&#xBA70; hepatocytes &#xD3EC;&#xD568; &#xD558; &#xC5EC; &#xBAA8;&#xB4E0; 3 &#xAC1C;&#xC758; &#xC138;&#xADE0; &#xCE35;&#xC758; &#xC138;&#xD3EC;&#xB85C; &#xBD84;&#xD654;. Humaninduced &#xB9CC;&#xB2A5; &#xC904;&#xAE30; &#xC138;&#xD3EC; (iPSCs) &#xD658;&#xC790;&#xC5D0;&#xC11C; &#xD30C;&#xC0DD; &#xB41C; &#xB610;&#xB294; &#xAC1C;&#xC778;&#xC758; &#xC140; hepatocyte &#xAC19;&#xC740; &#xC140; &#xC815;&#xC758; &#xB41C; &#xC720;&#xC804;&#xC790; &#xC131;&#xBD84;&#xACFC; &#xC0DD;&#xC131; &#xC0C8;&#xB85C;&#xC6B4; &#xAE30;&#xD68C;&#xB97C; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;. &#xC5EC;&#xAE30;, &#xC6B0;&#xB9AC;&#xAC00; HBV&#xC640; HCV&#xC5D0; &#xC0AC;&#xC6A9; &#xB418;&#xB294; &#xBAA8;&#xB378;&#xC758; &#xD604;&#xC7AC; &#xAD00;&#xC810;&#xC744; &#xAC80;&#xD1A0;&#xD560; &#xAC83;&#xC785;&#xB2C8;&#xB2E4; &#xACF5;&#xBD80;, &#xADF8;&#xB9AC;&#xACE0; &#xC778;&#xAC04;&#xC758; Ipscs&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC; &#xAC1C;&#xC778; &#xC124;&#xC815; &#xB41C; &#xB9C8;&#xC6B0;&#xC2A4; &#xBAA8;&#xB378;&#xC744; &#xC18C;&#xAC1C; &#xD569;&#xB2C8;&#xB2E4;. &#xC774; &#xC0C8;&#xB85C;&#xC6B4; &#xB9C8;&#xC6B0;&#xC2A4; &#xBAA8;&#xB378; HBV&#xC640; HCV hepatocytes&#xC758; &#xC790;&#xD654; &#xC728;&#xC5D0; &#xC720;&#xC804;&#xC758; &#xC601;&#xD5A5;&#xC744; &#xD504;&#xB85C; &#xBE59; &#xC778;&#xAC04;&#xC758; hepatocytes&#xC5D0; HBV&#xC640; HCV&#xC758; &#xC9C1;&#xC811; &#xC870;&#xC0AC;&#xB97C; &#xCD09;&#xC9C4;&#xD560; &#xAC83; &#xC774;&#xB2E4;.

Humanized murine &#xBAA8;&#xB378; HBV&#xC640; HCV&#xB294; &#xC778;&#xAC04; &#xC720;&#xB3C4; &#xB9CC;&#xB2A5; &#xC904;&#xAE30; &#xC138;&#xD3EC;&#xB97C; &#xC0AC;&#xC6A9; &#xD558; &#xC5EC;.

La infecci&#xF3;n de la hepatitis B (VHB) y hepatitis C (VHC) se traduce en resultados heterog&#xE9;neos desde una infecci&#xF3;n asintom&#xE1;tica a la infecci&#xF3;n aguda cr&#xF3;nica que conduce a la cirrosis y el carcinoma hepatocelular (CHC). En los modelos in vitro con hepatocitos humanos de los animales, l&#xED;neas celulares de HCC, o en modelos in vivo de ratones transg&#xE9;nicos han contribuido de manera inestimable a la comprensi&#xF3;n de la patog&#xE9;nesis de VHB y VHC. Un modelo de rat&#xF3;n humanizado hecha por la reconstituci&#xF3;n de hepatocitos primarios humanos en el h&#xED;gado del rat&#xF3;n inmunodeficiente proporciona una oportunidad novela experimental que simula el crecimiento in vivo de los hepatocitos humanos. El acceso limitado a los hepatocitos humanos primarios necesaria la b&#xFA;squeda de otras fuentes celulares, como c&#xE9;lulas madre pluripotentes. Las c&#xE9;lulas madre embrionarias (hESCs) tienen las caracter&#xED;sticas de auto-renovaci&#xF3;n y pluripotencialidad y se diferencian en c&#xE9;lulas de las tres capas germinales, incluyendo los hepatocitos. Humaninduced c&#xE9;lulas madre pluripotentes (iPSCs) derivados de la paciente o de las propias c&#xE9;lulas individuales ofrecen una nueva oportunidad para generar hepatocitos como las c&#xE9;lulas con la composici&#xF3;n definida gen&#xE9;tica. Aqu&#xED;, vamos a revisar la perspectiva actual de los modelos utilizados para el VHB y el VHC estudio, e introducir el modelo de rat&#xF3;n personalizada utilizando iPSCs humanos. Este modelo de rat&#xF3;n novela facilitar la investigaci&#xF3;n directa del VHB y el VHC en los hepatocitos humanos, as&#xED; como investigando la influencia gen&#xE9;tica en la susceptibilidad de los hepatocitos para el VHB y el VHC.

Modelo murino humanizado para el VHB y el VHC Uso Humanos c&#xE9;lulas madre pluripotentes inducidas

During acute or chronic hepatitis B virus (HBV) infection, the virus can invade the male reproductive system, pass through the blood-testis barrier and integrate into the germ line, resulting in abnormal spermatozoa. However, the pathway remains unclear. The asialoglycoprotein receptor (ASGR), a potential receptor for HBV, is mainly distributed in hepatocytes. We have examined the distribution of ASGR in human testis and found it in the seminiferous tubules and interstitial region but its enrichment in human testis is much lower than that in liver. By multiple immunoenzyme histochemistry staining, ASGR was precisely co-localized with vimentin (Sertoli cell marker) but not proliferating cell nuclear antigen (spermatogonial cell marker) in testis tissue. ASGR was expressed in human Leydig cells, stromal cells in the seminiferous tubules and Sertoli cells but seldom in spermatogonial cells. Therefore, ASGR could provide HBV with access to the luminal compartment of human testis. The mechanism by which HBV invades germ cells remains unknown.

Expression pattern of asialoglycoprotein receptor in human testis.

Stem cell-derived hepatocytes represent promising models to study human liver biology and disease. This concise review discusses the recent progresses in the field, with a focus on human liver disease, drug metabolism and virus infection.

Modeling human liver biology using stem cell-derived hepatocytes.

In this study, human embryonic stem cell-derived hepatocytes (hESC-Heps) were investigated for their ability to support hepatitis C virus (HCV) infection and replication. hESC-Heps were capable of supporting the full viral life cycle, including the release of infectious virions. Although supportive, hESC-Hep viral infection levels were not as great as those observed in Huh7 cells. We reasoned that innate immune responses in hESC-Heps may lead to the low level of infection and replication. Upon further investigation, we identified a strong type III interferon response in hESC-Heps that was triggered by HCV. Interestingly, specific inhibition of the JAK/STAT signaling pathway led to an increase in HCV infection and replication in hESC-Heps. Of note, the interferon response was not evident in Huh7 cells. In summary, we have established a robust cell-based system that allows the in-depth study of virus-host interactions in vitro.

Modulating innate immunity improves hepatitis C virus infection and replication in stem cell-derived hepatocytes.

To improve the efficiency, reproducibility and consistency of the PEI-based transfection method that is often used in preparation of recombinant lentiviral or retroviral vectors.

A rapid and efficient polyethylenimine-based transfection method to prepare lentiviral or retroviral vectors: useful for making iPS cells and transduction of primary cells.

To develop a rapid and accurate assay system for screening inhibitors or enhancing agents targeting the transactivation capability of hepatitis B virus X protein (HBx) that activates cellular promoters in host cells to facilitate viral replication.

An efficient rapid system for assaying HBx-mediated transactivation.

Polyethyleneimine (PEI), a cationic polymer vehicle, forms a complex with DNA which then can carry anionic nucleic acids into eukaryotic cells. PEI-based transfection is widely used for transient transfection of plasmid DNA. The efficiency of PEI-based transfection is affected by numerous factors, including the way the PEI/DNA complex is prepared, the ratio of PEI to DNA, the concentration of DNA, the storage conditions of PEI solutions, and more. Considering the major influencing factors, PEI-based transfection has been optimized to improve its efficiency, reproducibility, and consistency. This protocol outlines the steps for ordinary transient transfection and lentiviral production using PEI. © 2017 by John Wiley & Sons, Inc.

Optimized PEI-based Transfection Method for Transient Transfection and Lentiviral Production.

Hepatitis B virus (HBV) is a human hepatotropic virus. However, HBV infection also occurs at extrahepatic sites, but the relevant host factors required for HBV infection in non-hepatic cells are only partially understood. In this article, a non-hepatic cell culture model is constructed by exogenous expression of four host genes (NTCP, HNF4α, RXRα and PPARα) in human non-hepatic 293T cells. This cell culture model supports HBV entry, transcription and replication, as evidenced by the detection of HBV pgRNA, HBV cccDNA, HBsAg, HBeAg, HBcAg and HBVDNA. Our results suggest that the above cellular factors may play a key role in HBV infection of non-hepatic cells. This model will facilitate the identification of host genes that support extrahepatic HBV infection.

Defined host factors support HBV infection in non-hepatic 293T cells.

Hepatitis B virus (HBV) infection affects ~350 million people and poses a major public health problem worldwide. HBV is a major cause of cirrhosis and hepatocellular carcinoma. Fewer than 5% of HBV-infected adults (but up to 90% of HBV-infected infants and children) develop chronic HBV infection as indicated by continued, detectable expression of hepatitis B surface antigen (HBsAg) for at least 6 months after the initial infection. Increasing evidence indicates that HBV interacts with innate immunity signaling pathways of hepatocytes to suppress innate immunity. However, it is still not clear how HBV avoids monitoring by the innate immunity of hepatocytes and whether the innate immunity of hepatocytes can be effective against HBV if re-triggered. Moreover, a deep understanding of virus-host interactions is important in developing new therapeutic strategies for the treatment of HBV infection. In this review, we summarize the current knowledge regarding how HBV represses innate immune recognition, as well as recent progress with respect to in vitro models for studying HBV infection and innate immunity.

The Interactions between HBV and the Innate Immunity of Hepatocytes.

Hepatitis B virus (HBV) infection may affect sperm motility in patients with HBV. HBV surface protein (HBs) decreases mitochondrial membrane potential, impairs motility and induces apoptotic-like changes in human spermatozoa. However, little is known about how human spermatozoa respond to reactive oxygen species (ROS; mainly peroxides) induced by HBs. In this study, HBs induced supraphysiological ROS levels in human spermatozoa and reduced the formation of 2-cell embryos (obtained from hamster oocytes and human spermatozoa). HBs induced a pre-apoptotic status in human spermatozoa, as well as antioxidant defences by increasing glutathione peroxidase 4 (GPX4) and peroxiredoxin 5 (PRDX5) levels. These results highlight the molecular mechanism responsible for the oxidative stress in human spermatozoa exposed to HBV and the antioxidant defence response involving GPX4 and PRDX5.

ArticleTotal : 2	Year
Publication title Cited by 1	2020
Publication title Cited by 3	2021

Article	Year
Humanized murine model for HBV and HCV using human induced pluripotent stem cells. Archives of pharmacal research\| PubMed ID: 22370780	2012
Expression pattern of asialoglycoprotein receptor in human testis. Cell and tissue research\| PubMed ID: 23604802	2013
Modeling human liver biology using stem cell-derived hepatocytes. International journal of molecular sciences\| PubMed ID: 24201130	2013
Modulating innate immunity improves hepatitis C virus infection and replication in stem cell-derived hepatocytes. Stem cell reports\| PubMed ID: 25068132	2014
A rapid and efficient polyethylenimine-based transfection method to prepare lentiviral or retroviral vectors: useful for making iPS cells and transduction of primary cells. Biotechnology letters\| PubMed ID: 27193760	2016
An efficient rapid system for assaying HBx-mediated transactivation. Biotechnology letters\| PubMed ID: 28386736	2017
Optimized PEI-based Transfection Method for Transient Transfection and Lentiviral Production. Current protocols in chemical biology\| PubMed ID: 28910855	2017
Defined host factors support HBV infection in non-hepatic 293T cells. Journal of cellular and molecular medicine\| PubMed ID: 31930674	2020
The Interactions between HBV and the Innate Immunity of Hepatocytes. Viruses\| PubMed ID: 32151000	2020
Hepatitis B virus surface protein induces oxidative stress by increasing peroxides and inhibiting antioxidant defences in human spermatozoa. Reproduction, fertility, and development\| PubMed ID: 32998796	2020

Pingnan Sun

Biography

JoVE Journal Publications

Other Publications