creation-cardiac-tissue-exhibiting-mechanical-integration-spheroids

Creation of Cardiac Tissue Exhibiting Mechanical Integration of Spheroids Using 3D Bioprinting

This protocol describes 3D bioprinting of cardiac tissue without the use of biomaterials, using only cells. Cardiomyocytes, endothelial cells and ...

Johns Hopkins University

&#x4F5C;&#x4E3A;&#x539F;&#x578B;&#x6765;&#x6E90;&#x53EF;&#x4EE5;&#x5728;&#x836F;&#x7269;&#x53D1;&#x73B0;&#x548C;&#x7EC6;&#x80DE;&#x6CBB;&#x7597;&#x4E2D;&#x4F7F;&#x7528;&#x6D3E;&#x751F;&#x5FC3;&#x808C;&#x7EC6;&#x80DE;&#x51FA;&#x73B0;&#x4E86;&#x4EBA;&#x7C7B;&#x80DA;&#x80CE;&#x5E72;&#x7EC6;&#x80DE;&#x3002;&#x4F46;&#x662F;&#xFF0C;&#x8FD9;&#x79CD;&#x5E94;&#x7528;&#x7A0B;&#x5E8F;&#x9700;&#x8981;&#x8FD9;&#x4E9B;&#x5FC3;&#x808C;&#x7EC6;&#x80DE; (hESC CMs) &#x5FE0;&#x5B9E;&#x5730;&#x590D;&#x8FF0;&#x6210;&#x4EBA;&#x7EC6;&#x80DE;&#xFF0C;&#x5C24;&#x5176;&#x662F;&#x5728;&#x4ED6;&#x4EEC;&#x7684;&#x7535;&#x751F;&#x7406;&#x53CA;&#x6536;&#x7F29;&#x529F;&#x80FD;&#x7684;&#x751F;&#x7406;&#x3002;&#x6211;&#x4EEC;&#x68C0;&#x8BA8;&#x6240;&#x77E5;&#x7684; hESC CMs &#x7684;&#x8DF3;&#x52A8;&#x7387;&#x3001; &#x52A8;&#x4F5C;&#x7535;&#x4F4D;&#x7279;&#x5F81;&#x3001; &#x79BB;&#x5B50;&#x7535;&#x6D41;&#x548C;&#x8702;&#x7A9D;&#x8026;&#x5408;&#xFF0C;&#x4EE5;&#x53CA;&#x5176;&#x6536;&#x7F29;&#x7684;&#x9499;&#x7684;&#x7535;&#x751F;&#x7406;&#x5FAA;&#x73AF;&#x548C;&#x6536;&#x7F29;&#x3002;&#x6211;&#x4EEC;&#x8FD8;&#x8BA8;&#x8BBA;&#x5728;&#x7EC6;&#x80DE;&#x8868;&#x578B;&#x7684;&#x975E;&#x5747;&#x8D28;&#x6027;&#x4EA7;&#x751F;&#x53D8;&#x5F02;&#x6027;&#x7684;&#x5FC3;&#x810F;&#x5206;&#x5316;&#x3001; &#x6210;&#x719F;&#x548C;&#x6587;&#x5316;&#x6761;&#x4EF6;&#xFF0C;&#x5E76;&#x6C47;&#x603B;&#x5DF2;&#x5B9E;&#x65BD;&#x4EE5;&#x51CF;&#x5C11;&#x8FD9;&#x79CD;&#x5F02;&#x8D28;&#x6027;&#x7684;&#x73B0;&#x884C;&#x6218;&#x7565;&#x3002;&#x6700;&#x540E;&#xFF0C;&#x6211;&#x4EEC;&#x63D0;&#x51FA;&#x4ECE;&#x5149;&#x5B66;&#x5730;&#x56FE; hESC &#x5398;&#x7C73;&#x96C6;&#x7FA4;&#x7684;&#x539F;&#x7535;&#x751F;&#x7406;&#x6570;&#x636E;&#x3002;

&#x4ECE;&#x4EBA;&#x7C7B;&#x80DA;&#x80CE;&#x5E72;&#x7EC6;&#x80DE;&#x7684;&#x5FC3;&#x808C;&#x7EC6;&#x80DE;&#x7535;&#x751F;&#x7406;&#x548C;&#x6536;&#x7F29;&#x529F;&#x80FD;&#x3002;

Human embryonic stem cells have emerged as the prototypical source from which cardiomyocytes can be derived for use in drug discovery and cell therapy. However, such applications require that these cardiomyocytes (hESC-CMs) faithfully recapitulate the physiology of adult cells, especially in relation to their electrophysiological and contractile function. We review what is known about the electrophysiology of hESC-CMs in terms of beating rate, action potential characteristics, ionic currents, and cellular coupling as well as their contractility in terms of calcium cycling and contraction. We also discuss the heterogeneity in cellular phenotypes that arises from variability in cardiac differentiation, maturation, and culture conditions, and summarize present strategies that have been implemented to reduce this heterogeneity. Finally, we present original electrophysiological data from optical maps of hESC-CM clusters.

Electrophysiological and contractile function of cardiomyocytes derived from human embryonic stem cells.

Cellules souches embryonnaires humaines sont apparus comme la source prototypique, d&#39;o&ugrave; des cardiomyocytes peuvent &ecirc;tre d&eacute;riv&eacute;s destin&eacute;s &agrave; la pharmacoth&eacute;rapie de d&eacute;couverte et de la cellule. Toutefois, ces applications exigent que ces cardiomyocytes (CSEh-CMs) r&eacute;capituler fid&egrave;lement la physiologie des cellules adultes, en particulier en ce qui concerne leur fonction &eacute;lectrophysiologique et contractile. Nous passons en revue ce que l&#39;on sait de l&#39;&eacute;lectrophysiologie de CSEh-CMs en termes de battre le taux, caract&eacute;ristiques du potentiel d&#39;action, les courants ioniques et couplage cellulaire ainsi que leur contractilit&eacute; en termes de calcium cyclisme et contraction. Nous &eacute;galement discuter de l&#39;h&eacute;t&eacute;rog&eacute;n&eacute;it&eacute; dans les ph&eacute;notypes cellulaires qui r&eacute;sulte de la variabilit&eacute; des conditions de culture, la maturation et diff&eacute;renciation cardiaque et r&eacute;sumer les strat&eacute;gies actuelles qui ont &eacute;t&eacute; mis en place afin de r&eacute;duire cette h&eacute;t&eacute;rog&eacute;n&eacute;it&eacute;. Enfin, nous pr&eacute;sentons des donn&eacute;es &eacute;lectrophysiologiques originales de cartes optiques des grappes de CSEh-CM.

Fonction &eacute;lectrophysiologique et contractile des cardiomyocytes d&eacute;riv&eacute;s de cellules souches embryonnaires humaines.

Menschlicher embryonaler Stammzellen sind als prototypische Quelle entstanden aus denen Cardiomyocytes f&uuml;r den Einsatz in Droge-Entdeckung und Zelltherapie abgeleitet werden k&ouml;nnen. Solche Anwendungen erfordern jedoch, dass diese Cardiomyocytes (hESC-CMs) getreulich die Physiologie der adulten Zellen, insbesondere im Hinblick auf ihre Funktion elektrophysiologische und KONTRAKTILER rekapitulieren. Wir &uuml;berpr&uuml;fen, was &uuml;ber die Elektrophysiologie des hESC-CMs in Bezug auf die Bewertung, Aktionspotenzial Merkmale, Ionische Str&ouml;mungen, und zellul&auml;re Kupplung sowie ihrer Kontraktilit&auml;t in Bezug auf Kalzium zu schlagen bekannt ist Radfahren und Kontraktion. Wir auch diskutieren die Heterogenit&auml;t in zellul&auml;ren Ph&auml;notypen, die durch die Variabilit&auml;t der kardialen Differenzierung, Reifung und Kulturbedingungen entsteht und Zusammenfassen der aktuellen Strategien, die umgesetzt wurden, um diese Heterogenit&auml;t zu reduzieren. Schlie&szlig;lich stellen wir Originale elektrophysiologische Daten aus optischen Karten hESC-CM-Cluster.

Elektrophysiologische und kontraktilen Funktion von Cardiomyocytes abgeleitet von humanen embryonalen Stammzellen.

Cellule staminali embrionali umane sono emersi come il prototipica fonte da cui cardiomyocytes pu&ograve; essere derivata per l&#39;uso in terapia farmacologica delle cellule e di scoperta. Tuttavia, tali applicazioni richiedono che questi cardiomyocytes (hESC-CMs) riassumere fedelmente la fisiologia delle cellule adulte, soprattutto in relazione alla loro funzione elettrofisiologica e contrattile. Passiamo in rassegna ci&ograve; che &egrave; noto circa l&#39;elettrofisiologia del hESC-CMs in termini battendo il tasso, le caratteristiche del potenziale d&#39;azione, correnti ioniche e accoppiamento cellulare cos&igrave; come loro contrattilit&agrave; in termini di calcio ciclismo e contrazione. Abbiamo anche discutere l&#39;eterogeneit&agrave; in fenotipi cellulari che nasce dalla variabilit&agrave; nelle condizioni di coltura, la maturazione e la differenziazione cardiache e riassumere le attuali strategie che sono state attuate per ridurre questa eterogeneit&agrave;. Infine, vi presentiamo i dati elettrofisiologici originali da mappe ottici di cluster hESC-CM.

Funzione contrattile ed elettrofisiologica di cardiomiociti derivati da cellule staminali embrionali umane.

&#x30D2;&#x30C8;&#x80DA;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x3001;&#x5FC3;&#x7B4B;&#x7D30;&#x80DE;&#x306E;&#x767A;&#x898B;&#x3068;&#x7D30;&#x80DE;&#x306E;&#x85AC;&#x7269;&#x7642;&#x6CD5;&#x3067;&#x4F7F;&#x7528;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x6D3E;&#x751F;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x30D7;&#x30ED;&#x30C8;&#x30BF;&#x30A4;&#x30D7;&#x306E;&#x30BD;&#x30FC;&#x30B9;&#x3068;&#x3057;&#x3066;&#x6D6E;&#x4E0A;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x305F;&#x3060;&#x3057;&#x3001;&#x3053;&#x306E;&#x3088;&#x3046;&#x306A;&#x30A2;&#x30D7;&#x30EA;&#x30B1;&#x30FC;&#x30B7;&#x30E7;&#x30F3;&#x306F;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x5FC3;&#x7B4B;&#x7D30;&#x80DE; (hESC-CMs) &#x304C;&#x5FE0;&#x5B9F;&#x306B;&#x96FB;&#x6C17;&#x751F;&#x7406;&#x5B66;&#x7684;&#x304A;&#x3088;&#x3073;&#x53CE;&#x7E2E;&#x6A5F;&#x80FD;&#x306B;&#x95A2;&#x9023;&#x3057;&#x3066;&#x7279;&#x306B;&#x3001;&#x5927;&#x4EBA;&#x306E;&#x7D30;&#x80DE;&#x306E;&#x751F;&#x7406;&#x5B66;&#x3092;&#x8981;&#x7D04;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3059;&#x3002;&#x6211;&#x3005; &#x306F;&#x306E;&#x305F;&#x3081;&#x306B;&#x60AA;&#x540D;&#x9AD8;&#x304F; CMs &#x7387;&#x3001;&#x6D3B;&#x52D5;&#x96FB;&#x4F4D;&#x7279;&#x6027;&#x3001;&#x30A4;&#x30AA;&#x30F3;&#x96FB;&#x6D41;&#x3068;&#x7D30;&#x80DE;&#x7D50;&#x5408;&#x3001;&#x30AB;&#x30EB;&#x30B7;&#x30A6;&#x30E0;&#x306E;&#x9762;&#x3067;&#x5F7C;&#x3089;&#x306E;&#x53CE;&#x7E2E;&#x3092;&#x7834;&#x3063;&#x3066;&#x306E;&#x9762;&#x3067;&#x306E;&#x96FB;&#x6C17;&#x751F;&#x7406;&#x5B66;&#x306B;&#x95A2;&#x3059;&#x308B;&#x65E2;&#x77E5;&#x306E;&#x4E8B;&#x67C4;&#x3092;&#x78BA;&#x8A8D;&#x30B5;&#x30A4;&#x30AF;&#x30EA;&#x30F3;&#x30B0;&#x3068;&#x53CE;&#x7E2E;&#x3002;&#x6211;&#x3005; &#x3082;&#x5FC3;&#x81D3;&#x5206;&#x5316;&#x3001;&#x6210;&#x719F;&#x304A;&#x3088;&#x3073;&#x57F9;&#x990A;&#x6761;&#x4EF6;&#x306E;&#x5909;&#x52D5;&#x304B;&#x3089;&#x751F;&#x3058;&#x308B;&#x7D30;&#x80DE;&#x8868;&#x73FE;&#x578B;&#x306E;&#x4E0D;&#x5747;&#x8CEA;&#x3092;&#x8AAC;&#x660E;&#x3057;&#x3001;&#x3053;&#x306E;&#x4E0D;&#x5747;&#x4E00;&#x6027;&#x3092;&#x6E1B;&#x3089;&#x3059;&#x305F;&#x3081;&#x306B;&#x5B9F;&#x88C5;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x73FE;&#x5728;&#x306E;&#x6226;&#x7565;&#x3092;&#x307E;&#x3068;&#x3081;&#x307E;&#x3059;&#x3002;&#x6700;&#x5F8C;&#x306B;&#x3001;&#x5149;&#x306E;&#x30DE;&#x30C3;&#x30D7;&#x306E;&#x305F;&#x3081;&#x306B;&#x60AA;&#x540D;&#x9AD8;&#x304F; CM &#x30AF;&#x30E9;&#x30B9;&#x30BF;&#x30FC;&#x304B;&#x3089;&#x5143;&#x306E;&#x96FB;&#x6C17;&#x751F;&#x7406;&#x5B66;&#x7684;&#x30C7;&#x30FC;&#x30BF;&#x3092;&#x63D0;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;

&#x4EBA;&#x9593;&#x306E;&#x80DA;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x6D3E;&#x751F;&#x3057;&#x305F;&#x5FC3;&#x7B4B;&#x7D30;&#x80DE;&#x306E;&#x96FB;&#x6C17;&#x751F;&#x7406;&#x5B66;&#x7684;&#x304A;&#x3088;&#x3073;&#x53CE;&#x7E2E;&#x6A5F;&#x80FD;&#x3002;

&#xC778;&#xAC04; &#xBC30;&#xC544; &#xC904;&#xAE30; &#xC138;&#xD3EC;&#xB294; cardiomyocytes &#xC57D;&#xBB3C; &#xBC1C;&#xACAC; &#xBC0F; &#xC138;&#xD3EC; &#xCE58;&#xB8CC;&#xC5D0; &#xC0AC;&#xC6A9; &#xD558;&#xAE30; &#xC704;&#xD574; &#xD30C;&#xC0DD; &#xB420; &#xC218; &#xC788;&#xB294; &#xC6D0;&#xD615; &#xC18C;&#xC2A4;&#xB3C4; &#xB5A0;&#xC624;&#xB974;&#xACE0; &#xC788;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;, &#xC774;&#xB7EC;&#xD55C; &#xC751;&#xC6A9; &#xD504;&#xB85C;&#xADF8;&#xB7A8;&#xC5D0;&#xC11C;&#xB294; &#xC774;&#xB7EC;&#xD55C; cardiomyocytes (hESC-CMs) &#xCDA9;&#xC2E4; &#xD558; &#xAC8C; &#xC790;&#xC2E0;&#xC758; electrophysiological &#xBC0F; &#xC218;&#xCD95; &#xC131; &#xAE30;&#xB2A5;&#xC5D0; &#xAD00;&#xD558;&#xC5EC; &#xD2B9;&#xD788; &#xC131;&#xC778; &#xC138;&#xD3EC; &#xC0DD;&#xB9AC;&#xD559; &#xC815;&#xB9AC;. &#xC6B0;&#xB9AC;&#xAC00; &#xAC80;&#xD1A0; &#xD558;&#xB294; &#xBC15;&#xB3D9; &#xC18D;&#xB3C4;, &#xD65C;&#xB3D9; &#xC804;&#xC704; &#xD2B9;&#xC131;, &#xC774;&#xC628; &#xC804;&#xB958; &#xBC0F; &#xC138;&#xD3EC; &#xACB0;&#xD569; &#xBFD0;&#xB9CC; &#xC544;&#xB2C8;&#xB77C; &#xCE7C;&#xC298;&#xC758; &#xC810;&#xC5D0;&#xC11C; &#xADF8;&#xB4E4;&#xC758; &#xC218;&#xCD95; &#xBA74;&#xC5D0;&#xC11C; hESC-Cms&#xC758; &#xC804;&#xAE30; &#xC0DD;&#xB9AC;&#xD559;&#xC5D0; &#xB300; &#xD55C; &#xC54C;&#xB824;&#xC9C4; &#xC0AC;&#xC774;&#xD074;&#xB9C1; &#xBC0F; &#xC218;&#xCD95;. &#xB610;&#xD55C; &#xC2EC;&#xC7A5; &#xBD84;&#xD654;, &#xC131;&#xC219;, &#xADF8;&#xB9AC;&#xACE0; &#xBB38;&#xD654; &#xC870;&#xAC74;&#xC5D0; &#xC788;&#xB294; &#xBCC0;&#xD654;&#xC5D0;&#xC11C; &#xBC1C;&#xC0DD; &#xD558;&#xB294; &#xC140;&#xB8F0;&#xB7EC; &#xACE0;&#xAE30;&#xAC00; &#xC9C8;&#xC758; &#xC124;&#xBA85; &#xD558; &#xACE0;&#xC774; &#xC904;&#xC774;&#xAE30; &#xC704;&#xD574; &#xAD6C;&#xD604; &#xB41C; &#xD604;&#xC7AC;&#xC758; &#xC804;&#xB7B5;&#xC744; &#xC694;&#xC57D;. &#xB9C8;&#xC9C0;&#xB9C9;&#xC73C;&#xB85C;, &#xC6B0;&#xB9AC;&#xB294; hESC CM &#xD074;&#xB7EC;&#xC2A4;&#xD130;&#xC758; &#xAD11;&#xD559; &#xC9C0;&#xB3C4;&#xC5D0;&#xC11C; &#xC6D0;&#xB798; electrophysiological &#xB370;&#xC774;&#xD130; &#xD604;&#xC7AC;.

&#xC778;&#xAC04; &#xBC30;&#xC544; &#xC904;&#xAE30; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xD30C;&#xC0DD; &#xB41C; cardiomyocytes electrophysiological &#xBC0F; &#xC218;&#xCD95; &#xC131; &#xAE30;&#xB2A5;.

C&eacute;lulas madre embrionarias humanas han surgido como fuente protot&iacute;pica de que cardiomiocitos pueden ser derivados para su uso en el tratamiento farmacol&oacute;gico de descubrimiento y de la c&eacute;lula. Sin embargo, estas aplicaciones requieren que estos cardiomiocitos (hESCs-CMs) recapitulan fielmente la fisiolog&iacute;a de las c&eacute;lulas adultas, especialmente en relaci&oacute;n con su funci&oacute;n contr&aacute;ctil y electrofisiol&oacute;gico. Repasamos lo que se conoce sobre la electrofisiolog&iacute;a de hESC-CMs en t&eacute;rminos de vencer a tarifa, caracter&iacute;sticas del potencial de acci&oacute;n, las corrientes i&oacute;nicas y acoplamiento celular as&iacute; como su contractilidad en t&eacute;rminos de calcio ciclismo y contracci&oacute;n. Tambi&eacute;n discutimos la heterogeneidad en los fenotipos celulares que surge de la variabilidad cardiaca diferenciaci&oacute;n, maduraci&oacute;n y condiciones de cultivo y resumir las estrategias actuales que se han implementado para reducir esta heterogeneidad. Finalmente, presentamos los datos electrofisiol&oacute;gicos originales de mapas &oacute;pticos de racimos de c&eacute;lulas-CM.

Funci&oacute;n contr&aacute;ctil y electrofisiol&oacute;gica de cardiomiocitos derivadas de c&eacute;lulas madre embrionarias humanas.

&#x81EA;&#x4EBA;&#x7C7B;&#x8BF1;&#x5BFC;&#x7684;&#x591A;&#x529F;&#x80FD;&#x5E72;&#x7EC6;&#x80DE;&#x5206;&#x5316;&#x5FC3;&#x808C;&#x7EC6;&#x80DE; (hiPSC-CMs) &#x7684;&#x7B2C;&#x4E00;&#x4E2A;&#x8BF4;&#x660E;&#xFF0C;&#x8FD9;&#x4E9B;&#x7EC6;&#x80DE;&#x83B7;&#x5F97;&#x4E86;&#x83AB;&#x5927;&#x7684;&#x5174;&#x8DA3;&#xFF0C;&#x5176;&#x6F5C;&#x5728;&#x4F7F;&#x7528;&#x75C5;&#x4EBA;&#x5177;&#x4F53;&#x5206;&#x6790;&#x548C;&#x6CBB;&#x7597;&#x3002;&#x6B64;&#x5916;&#xFF0C;hiPSC CMs &#x53EF;&#x4EE5;&#x6765;&#x81EA;&#x6350;&#x8D60;&#x8005;&#x4ECE;&#x7279;&#x5B9A;&#x7684;&#x5FC3;&#x810F;&#x75BE;&#x75C5;&#xFF0C;&#x4F7F;&#x4F53;&#x5916;&#x4EBA;&#x7C7B;&#x75BE;&#x75C5;&#x6A21;&#x578B;&#x7684;&#x673A;&#x7406;&#x7814;&#x7A76;&#x548C;&#x6CBB;&#x7597;&#x836F;&#x7269;&#x8BC4;&#x4F30;&#x60A3;&#x8005;&#x7684;&#x7EC6;&#x80DE;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x5176;&#x7535;&#x751F;&#x7406;&#x548C;&#x6536;&#x7F29;&#x5168;&#x9762;&#x7684;&#x4E86;&#x89E3;&#x662F;&#x529F;&#x80FD;&#x7684;&#x5FC5;&#x8981;&#x4E4B;&#x524D;&#x53EF;&#x4EE5;&#x5B9E;&#x73B0;&#x8FD9;&#x4E00;&#x6F5C;&#x529B;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#xFF0C;&#x6211;&#x4EEC;&#x5BA1;&#x67E5;&#x8FD9;&#x4E00;&#x65B0;&#x5174;&#x9886;&#x57DF;&#x4ECE;&#x529F;&#x80FD;&#x7684;&#x89D2;&#x5EA6;&#xFF0C;&#x7279;&#x522B;&#x4FA7;&#x91CD;&#x4E8E;&#x6253;&#x7387;&#x3001; &#x52A8;&#x4F5C;&#x7535;&#x4F4D;&#x3001; &#x79BB;&#x5B50;&#x7535;&#x6D41;&#x3001; &#x591A;&#x7EC6;&#x80DE;&#x4F20;&#x5BFC;&#x3001; &#x9499;&#x5904;&#x7406;&#x548C;&#x6536;&#x7F29;&#x3002;&#x6211;&#x4EEC;&#x8FDB;&#x4E00;&#x6B65;&#x603B;&#x7ED3;&#x9057;&#x4F20;&#x6027;&#x5FC3;&#x808C;&#x75BE;&#x75C5;&#x7684;&#x5BA1;&#x67E5;&#x73B0;&#x5B58; hiPSC &#x5398;&#x7C73;&#x75BE;&#x75C5;&#x6A21;&#x578B;&#x3002;

&#x5FC3;&#x808C;&#x7EC6;&#x80DE;&#x6765;&#x81EA;&#x4EBA;&#x7C7B;&#x8BF1;&#x5BFC;&#x591A;&#x80FD;&#x5E72;&#x7EC6;&#x80DE;&#x4F5C;&#x4E3A;&#x6B63;&#x5E38;&#x53CA;&#x60A3;&#x75C5;&#x5FC3;&#x810F;&#x7535;&#x751F;&#x7406;&#x548C;&#x6536;&#x7F29;&#x6027;&#x7684;&#x6A21;&#x5F0F;&#x3002;

Since the first description of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), these cells have garnered tremendous interest for their potential use in patient-specific analysis and therapy. Additionally, hiPSC-CMs can be derived from donor cells from patients with specific cardiac disorders, enabling inÂ vitro human disease models for mechanistic study and therapeutic drug assessment. However, a full understanding of their electrophysiological and contractile function is necessary before this potential can be realized. Here, we review this emerging field from a functional perspective, with particular emphasis on beating rate, action potential, ionic currents, multicellular conduction, calcium handling and contraction. We further review extant hiPSC-CM disease models that recapitulate genetic myocardial disease.

Cardiomyocytes derived from human induced pluripotent stem cells as models for normal and diseased cardiac electrophysiology and contractility.

Depuis la premi&egrave;re description des cardiomyocytes d&eacute;riv&eacute;s de cellules souches humaines pluripotentes induites (hiPSC-CMs), ces cellules ont valu un int&eacute;r&ecirc;t &eacute;norme pour leur utilisation potentielle dans l&#39;analyse propre au patient et de la th&eacute;rapie. En outre, hiPSC-CMs peuvent &ecirc;tre d&eacute;riv&eacute;s de cellules provenant de patients souffrant de troubles cardiaques sp&eacute;cifiques, permettant &agrave; des mod&egrave;les de maladies humaines in vitro pour &eacute;tude m&eacute;canistique et &eacute;valuation th&eacute;rapeutique. Toutefois, une pleine compr&eacute;hension de leur fonction &eacute;lectrophysiologique et contractile est n&eacute;cessaire avant que ce potentiel peut &ecirc;tre r&eacute;alis&eacute;. Ici, nous passons en revue ce domaine &eacute;mergent d&#39;un point de vue fonctionnel, avec un accent particulier sur le battant taux, potentiel d&#39;action, les courants ioniques, conduction multicellulaire, manipulation de calcium et contraction. Nous autres mod&egrave;les d&#39;examen existants hiPSC-CM maladie qui r&eacute;capitulent g&eacute;n&eacute;tique maladie myocardique.

Cardiomyocytes d&eacute;riv&eacute;s de cellules souches humaines pluripotentes induites comme des mod&egrave;les normaux et malade &eacute;lectrophysiologie cardiaque et la contractilit&eacute;.

Seit der ersten Beschreibung der menschlichen induzierten pluripotenten Stammzellen Cell-derived Cardiomyocytes (HiPSC-CMs) haben diese Zellen enormes Interesse f&uuml;r ihre m&ouml;gliche Verwendung in der Patienten-spezifische Analyse und Therapie vereinen. Dar&uuml;ber hinaus k&ouml;nnen HiPSC-CMs von Spenderzellen von Patienten mit bestimmten kardialen Erkrankungen, Aktivieren von in-vitro menschliche Krankheitsmodelle f&uuml;r mechanistische Untersuchung und therapeutisches Medikament Bewertung abgeleitet werden. Allerdings ist ein umfassendes Verst&auml;ndnis ihrer elektrophysiologische und kontraktilen Funktion notwendig, bevor dieses Potenzial realisiert werden kann. Hier schauen wir uns diese aufstrebenden Gebiet aus einer funktionalen Perspektive, mit besonderem Schwerpunkt auf Preise, Aktionspotential, Ionische Str&ouml;mungen, vielzelligen Leitverm&ouml;gen, Kalzium-Handhabung und Kontraktion zu schlagen. Wir weitere &Uuml;berpr&uuml;fung erhaltenen HiPSC-CM-Krankheitsmodelle, die myokardialen Erbkrankheit zu rekapitulieren.

Cardiomyocytes abgeleitet von menschlichen induzierten pluripotenten Stammzellen als Modelle f&uuml;r normalen und Kranken kardiale Elektrophysiologie und Kontraktilit&auml;t.

Poich&eacute; la prima descrizione di cardiomiociti derivati da cellule staminali umane pluripotenti indotte (hiPSC-CMs), queste cellule hanno raccolto enorme interesse per il loro uso potenziale paziente-specifici analisi e terapia. Inoltre, hiPSC-CMs pu&ograve; essere derivata dalle cellule del donatore da pazienti con disturbi cardiaci specifici, consentendo di modelli di malattie umane in vitro per Studi meccanicistici e valutazione terapeutica del farmaco. Tuttavia, una piena comprensione della loro funzione elettrofisiologica e contrattile &egrave; necessaria prima di questo potenziale pu&ograve; essere realizzato. Qui, passiamo in rassegna questo campo emergente dal punto di vista funzionale, con particolare enfasi su come battere il tasso, potenziale d&#39;azione, correnti ioniche, conduzione pluricellulari, calcio e contrazione. Abbiamo ulteriori modelli di malattia esistenti hiPSC-CM recensione che riassumere la malattia genetica del miocardio.

Cardiomiociti derivano da cellule staminali pluripotenti indotte umano come modelli per elettrofisiologia cardiaca normale e malato e la contrattilit&agrave;.

&#x3072;&#x3068;&#x8A98;&#x5C0E;&#x591A;&#x80FD;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#x7531;&#x6765;&#x5FC3;&#x7B4B;&#x7D30;&#x80DE; &#xFF08;&#x304B;&#x3089;&#x4F5C;&#x88FD;&#x3057;&#x305F; CMs&#xFF09; &#x306E;&#x6700;&#x521D;&#x306E;&#x8AAC;&#x660E;&#x306A;&#x306E;&#x3067;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x7D30;&#x80DE;&#x306F;&#x60A3;&#x8005;&#x56FA;&#x6709;&#x306E;&#x89E3;&#x6790;&#x3068;&#x6CBB;&#x7642;&#x306E;&#x6F5C;&#x5728;&#x7684;&#x306A;&#x4F7F;&#x7528;&#x306E;&#x305F;&#x3081;&#x306E;&#x9014;&#x65B9;&#x3082;&#x306A;&#x3044;&#x8208;&#x5473;&#x3092;&#x96C6;&#x3081;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x307E;&#x305F;&#x3001;&#x30C9;&#x30CA;&#x30FC;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x7279;&#x5B9A;&#x5FC3;&#x7B4B;&#x75C7;&#x306E;&#x60A3;&#x8005;&#x306E;&#x89E3;&#x660E;&#x3068;&#x6CBB;&#x7642;&#x85AC;&#x8A55;&#x4FA1;&#x306E;&#x305F;&#x3081;&#x306E; in vitro &#x3067;&#x30D2;&#x30C8;&#x75BE;&#x60A3;&#x30E2;&#x30C7;&#x30EB;&#x3092;&#x6709;&#x52B9;&#x306B;&#x3059;&#x308B;&#x3092;&#x304B;&#x3089;&#x4F5C;&#x88FD;&#x3057;&#x305F; CMs &#x7531;&#x6765;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x3002;&#x305F;&#x3060;&#x3057;&#x3001;&#x3053;&#x306E;&#x53EF;&#x80FD;&#x6027;&#x3092;&#x5B9F;&#x73FE;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x3067;&#x304D;&#x307E;&#x3059;&#x524D;&#x306B;&#x3001;&#x96FB;&#x6C17;&#x751F;&#x7406;&#x5B66;&#x7684;&#x304A;&#x3088;&#x3073;&#x53CE;&#x7E2E;&#x6A5F;&#x80FD;&#x306E;&#x5B8C;&#x5168;&#x306A;&#x7406;&#x89E3;&#x304C;&#x5FC5;&#x8981;&#x3067;&#x3059;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;&#x3053;&#x306E;&#x7387;&#x3001;&#x6D3B;&#x52D5;&#x96FB;&#x4F4D;&#x3001;&#x30A4;&#x30AA;&#x30F3;&#x96FB;&#x6D41;&#x3001;&#x591A;&#x7D30;&#x80DE;&#x306E;&#x4F1D;&#x5C0E;&#x3001;&#x30AB;&#x30EB;&#x30B7;&#x30A6;&#x30E0;&#x51E6;&#x7406;&#x3068;&#x53CE;&#x7E2E;&#x3092;&#x6253;&#x3064;&#x3053;&#x3068;&#x306B;&#x7279;&#x306B;&#x91CD;&#x70B9;&#x3092;&#x30D5;&#x30A1;&#x30F3;&#x30AF;&#x30B7;&#x30E7;&#x30CA;&#x30EB;&#x306A;&#x8996;&#x70B9;&#x304B;&#x3089;&#x306E;&#x65B0;&#x305F;&#x306A;&#x30D5;&#x30A3;&#x30FC;&#x30EB;&#x30C9;&#x3092;&#x78BA;&#x8A8D;&#x3057;&#x307E;&#x3059;&#x3002;&#x6211;&#x3005; &#x306F;&#x907A;&#x4F1D;&#x7684;&#x5FC3;&#x7B4B;&#x75BE;&#x60A3;&#x3092;&#x8981;&#x7D04;&#x30EC;&#x30D3;&#x30E5;&#x30FC;&#x73FE;&#x5B58;&#x3059;&#x308B;&#x304B;&#x3089;&#x4F5C;&#x88FD;&#x3057;&#x305F; CM &#x75BE;&#x60A3;&#x30E2;&#x30C7;&#x30EB;&#x3092;&#x3055;&#x3089;&#x306B;&#x3002;

&#x5FC3;&#x7B4B;&#x7D30;&#x80DE;&#x306F;&#x6B63;&#x5E38;&#x3068;&#x75C5;&#x6C17;&#x5FC3;&#x81D3;&#x96FB;&#x6C17;&#x751F;&#x7406;&#x5B66;&#x3068;&#x53CE;&#x7E2E;&#x306E;&#x305F;&#x3081;&#x306E;&#x30E2;&#x30C7;&#x30EB;&#x3068;&#x3057;&#x3066;&#x3072;&#x3068;&#x8A98;&#x5C0E;&#x591A;&#x80FD;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#x304B;&#x3089;&#x6D3E;&#x751F;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;

&#xC778;&#xAC04; &#xC720;&#xB3C4; &#xB9CC;&#xB2A5; &#xC904;&#xAE30; &#xC138;&#xD3EC; &#xC720;&#xB798; cardiomyocytes (hiPSC-CMs)&#xC758; &#xCCAB; &#xBC88;&#xC9F8; &#xC124;&#xBA85; &#xC774;&#xD6C4; &#xC774;&#xB7EC;&#xD55C; &#xC138;&#xD3EC; &#xD658;&#xC790; &#xAD00;&#xB828; &#xBD84;&#xC11D; &#xBC0F; &#xCE58;&#xB8CC;&#xC5D0; &#xB300; &#xD55C; &#xADF8;&#xB4E4;&#xC758; &#xC7A0;&#xC7AC;&#xC801;&#xC778; &#xC0AC;&#xC6A9;&#xC5D0; &#xB300; &#xD55C; &#xC5C4;&#xCCAD;&#xB09C; &#xAD00;&#xC2EC;&#xC744; &#xC5BB;&#xACE0; &#xC788;&#xB2E4;. &#xB610;&#xD55C;, hiPSC CMs &#xAE30;&#xACC4; &#xC5F0;&#xAD6C; &#xBC0F; &#xCE58;&#xB8CC; &#xC57D;&#xBB3C; &#xD3C9;&#xAC00; &#xB300; &#xD55C; &#xC0DD;&#xCCB4; &#xC678;&#xC5D0;&#xC11C; &#xC778;&#xAC04; &#xC9C8;&#xBCD1; &#xBAA8;&#xB378;&#xC744; &#xC0AC;&#xC6A9; &#xD558;&#xBA74; &#xD2B9;&#xC815; &#xC2EC;&#xC7A5; &#xC7A5;&#xC560;&#xB97C; &#xAC00;&#xC9C4; &#xD658;&#xC790;&#xC5D0;&#xC11C; &#xAE30;&#xC99D;&#xC790; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xD30C;&#xC0DD; &#xB420; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;,&#xC774; &#xC7A0;&#xC7AC;&#xB825;&#xC744; &#xC2E4;&#xD604; &#xD558;&#xAE30; &#xC804;&#xC5D0; electrophysiological &#xBC0F; &#xC218;&#xCD95; &#xAE30;&#xB2A5; &#xB4E4;&#xC758; &#xC644;&#xC804; &#xD55C; &#xC774;&#xD574;&#xB294; &#xD544;&#xC694;. &#xC5EC;&#xAE30;&#xC5D0;, &#xC6B0;&#xB9AC;&#xB294; &#xC18D;&#xB3C4;, &#xD65C;&#xB3D9; &#xC804;&#xC704;, &#xC774;&#xC628; &#xC804;&#xB958;, &#xB2E4;&#xC138;&#xD3EC; &#xC804;&#xB3C4;, &#xCE7C;&#xC298; &#xCC98;&#xB9AC; &#xBC0F; &#xC218;&#xCD95;&#xC5D0; &#xD2B9;&#xD788; &#xC911;&#xC810;&#xC744; &#xAE30;&#xB2A5;&#xC801; &#xAD00;&#xC810;&#xC5D0;&#xC11C;&#xC774; &#xC2E0;&#xD765; &#xD544;&#xB4DC;&#xB97C; &#xAC80;&#xD1A0; &#xD569;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC; &#xC720;&#xC804;&#xC790; &#xC2EC;&#xADFC; &#xC9C8;&#xD658; &#xC815;&#xB9AC; &#xAC80;&#xD1A0; &#xD604;&#xC874; hiPSC CM &#xC9C8;&#xBCD1; &#xBAA8;&#xB378; &#xCD94;&#xAC00;.

Cardiomyocytes&#xB294; &#xC815;&#xC0C1;&#xC801;&#xC774; &#xACE0; &#xBCD1;&#xC5D0; &#xAC78;&#xB9B0; &#xC2EC;&#xC7A5; &#xC804;&#xAE30; &#xC0DD;&#xB9AC;&#xD559; &#xBC0F; &#xC218;&#xCD95;&#xC5D0; &#xB300; &#xD55C; &#xBAA8;&#xB378;&#xB85C; &#xC778;&#xAC04; &#xC720;&#xB3C4; &#xB9CC;&#xB2A5; &#xC904;&#xAE30; &#xC138;&#xD3EC;&#xC5D0;&#xC11C; &#xD30C;&#xC0DD; &#xB41C;.

Desde la primera descripci&oacute;n de cardiomiocitos de derivados de c&eacute;lulas madre pluripotentes inducidas humanas (hiPSC-CMs), estas c&eacute;lulas han cosechado gran inter&eacute;s para su uso potencial en an&aacute;lisis espec&iacute;fico para cada paciente y la terapia. Adem&aacute;s, hiPSC-CMs pueden derivarse de las c&eacute;lulas del donante de pacientes con trastornos card&iacute;acos espec&iacute;ficos, permitiendo a modelos de enfermedades humanas in vitro para estudio mecanicista y evaluaci&oacute;n de drogas terap&eacute;uticas. Sin embargo, una comprensi&oacute;n completa de su funci&oacute;n contr&aacute;ctil y electrofisiol&oacute;gico es necesaria antes de que este potencial puede ser observada. Aqu&iacute; repasamos este campo emergente desde una perspectiva funcional, con especial &eacute;nfasis en vencer a tarifa, potencial de acci&oacute;n, las corrientes i&oacute;nicas, conducci&oacute;n multicelular, manejo del calcio y contracci&oacute;n. Tenemos m&aacute;s modelos de revisi&oacute;n existentes hiPSC-CM enfermedades que recapitulan enfermedad gen&eacute;tica del miocardio.

Cardiomiocitos derivan de c&eacute;lulas madre pluripotentes inducidas humanas como modelos para Electrofisiolog&iacute;a card&iacute;aca normal y enferma y la contractilidad.

The electrophysiological consequences of cardiomyocyte and myofibroblast interactions remain unclear, and the contribution of mechanical coupling between these two cell types is still poorly understood. In this study, we examined the time course and mechanisms by which addition of myofibroblasts activated by transforming growth factor-beta (TGF-β) influence the conduction velocity (CV) of neonatal rat ventricular cell monolayers. We observed that myofibroblasts affected CV within 30 min of contact and that these effects were temporally correlated with membrane deformation of cardiomyocytes by the myofibroblasts. Expression of dominant negative RhoA in the myofibroblasts impaired both myofibroblast contraction and myofibroblast-induced slowing of cardiac conduction, whereas overexpression of constitutive RhoA had little effect. To determine the importance of mechanical coupling between these cell types, we examined the expression of the two primary cadherins in the heart (N- and OB-cadherin) at cell-cell contacts formed between myofibroblasts and cardiomyocytes. Although OB-cadherin was frequently found at myofibroblast-myofibroblast contacts, very little expression was observed at myofibroblast-cardiomyocyte contacts. The myofibroblast-induced slowing of cardiac conduction was not prevented by silencing of OB-cadherin in the myofibroblasts, and could be reversed by inhibitors of mechanosensitive channels (gadolinium or streptomycin) and cellular contraction (blebbistatin). In contrast, N-cadherin expression was commonly observed at myofibroblast-cardiomyocyte contacts, and silencing of N-cadherin in myofibroblasts prevented the myofibroblast-dependent slowing of cardiac conduction. We propose that myofibroblasts can impair the electrophysiological function of cardiac tissue through the application of contractile force to the cardiomyocyte membrane via N-cadherin junctions.

Acute slowing of cardiac conduction in response to myofibroblast coupling to cardiomyocytes through N-cadherin.

Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are the most promising source of cardiomyocytes (CMs) for experimental and clinical applications, but their use is largely limited by a structurally and functionally immature phenotype that most closely resembles embryonic or fetal heart cells. The application of physical stimuli to influence hPSC-CMs through mechanical and bioelectrical transduction offers a powerful strategy for promoting more developmentally mature CMs. Here we summarize the major events associated with in vivo heart maturation and structural development. We then review the developmental state of in vitro derived hPSC-CMs, while focusing on physical (electrical and mechanical) stimuli and contributory (metabolic and hypertrophic) factors that are actively involved in structural and functional adaptations of hPSC-CMs. Finally, we highlight areas for possible future investigation that should provide a better understanding of how physical stimuli may promote in vitro development and lead to mechanistic insights. Advances in the use of physical stimuli to promote developmental maturation will be required to overcome current limitations and significantly advance research of hPSC-CMs for cardiac disease modeling, in vitro drug screening, cardiotoxicity analysis and therapeutic applications.

Physical developmental cues for the maturation of human pluripotent stem cell-derived cardiomyocytes.

A major consideration in the design of engineered cardiac tissues for the faithful representation of physiological behavior is the recapitulation of the complex topography and biochemistry of native tissue. In this study we present engineered heart slices (EHS), which consist of neonatal rat ventricular cells (NRVCs) seeded onto thin slices of decellularized cardiac tissue that retain important aspects of native extracellular matrix (ECM). To form EHS, rat or pig ventricular tissue was sectioned into 300 μm-thick, 5 to 16 mm-diameter disks, which were subsequently decellularized using detergents, spread on coverslips, and seeded with NRVCs. The organized fiber structure of the ECM remained after decellularization and promoted cell elongation and alignment, resulting in an anisotropic, functional tissue that could be electrically paced. Contraction decreased at higher pacing rates, and optical mapping revealed electrical conduction that was anisotropic with a ratio of approximately 2.0, rate-dependent shortening of the action potential and slowing of conduction, and slowing of conduction by the sodium channel blocker lidocaine. Reentrant arrhythmias could also be pace-induced and terminated. EHS constitute an attractive in vitro cardiac tissue in which cardiac cells are cultured on thin slices of decellularized cardiac ECM that provide important biochemical, structural, and mechanical cues absent in traditional cell cultures.

Engineered heart slices for electrophysiological and contractile studies.

Cardiac tissue engineering approaches have the potential to regenerate functional myocardium with intrinsic vascular networks. This study compared the relative effects of human adipose-derived stem/stromal cells (hASCs) and human dermal fibroblasts (hDFs) in cocultures with neonatal rat ventricular cardiomyocytes (NRVCMs) and human umbilical vein endothelial cells (HUVECs). At the same ratios of NRVCM:hASC and NRVCM:hDF, the hASC cocultures displayed shorter action potentials and maintained capture at faster pacing rates. Similarly, in coculture with HUVECs, hASC:HUVEC exhibited superior ability to support vascular capillary network formation relative to hDF:HUVEC. Based on these studies, a range of suitable cell ratios were determined to develop a triculture system. Six seeding ratios of NRVCM:hASC:HUVEC were tested and it was found that a ratio of 500:50:25 cells (i.e. 250,000:25,000:12,500 cells/cm(2) ) resulted in the formation of robust vascular networks while retaining action potential durations and propagation similar to pure NRVCM cultures. Tricultures in this ratio exhibited an average conduction velocity of 20 ± 2 cm/s, action potential durations at 80% repolarization (APD80 ) and APD30 of 122 ± 5 ms and 59 ± 4 ms, respectively, and maximum capture rate of 7.4 ± 0.6 Hz. The NRVCM control groups had APD80 and APD30 of 120 ± 9 ms and 51 ± 5 ms, with a maximum capture rate of 7.3 ± 0.2 Hz. In summary, the combination of hASCs in the appropriate ratios with NRVCMs and HUVECs can facilitate the formation of densely vascularized cardiac tissues that appear not to impact the electrophysiological function of cardiomyocytes negatively. Copyright © 2017 John Wiley & Sons, Ltd.

Article	Year
Electrophysiological and contractile function of cardiomyocytes derived from human embryonic stem cells. Progress in biophysics and molecular biology\| PubMed ID: 22958937	2012
Cardiomyocytes derived from human induced pluripotent stem cells as models for normal and diseased cardiac electrophysiology and contractility. Progress in biophysics and molecular biology\| PubMed ID: 22971665	2012
Acute slowing of cardiac conduction in response to myofibroblast coupling to cardiomyocytes through N-cadherin. Journal of molecular and cellular cardiology\| PubMed ID: 24412534	2014
Physical developmental cues for the maturation of human pluripotent stem cell-derived cardiomyocytes. Stem cell research & therapy\| PubMed ID: 25688759	2014
Engineered heart slices for electrophysiological and contractile studies. Biomaterials\| PubMed ID: 25934457	2015
Adipose-derived perivascular mesenchymal stromal/stem cells promote functional vascular tissue engineering for cardiac regenerative purposes. Journal of tissue engineering and regenerative medicine\| PubMed ID: 28103423	2017

Adriana Blazeski

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