Alternative Cultures for Human Pluripotent Stem Cell Production, Maintenance, and Genetic Analysis

Human pluripotent stem cells (hPSCs) hold great promise for regenerative medicine and biopharmaceutical applications. Currently, optimal culture and ...

National Institutes of Health

The extracellular aggregation of amyloid beta (Abeta) peptides and the intracellular hyperphosphorylation of tau at specific epitopes are pathological hallmarks of neurodegenerative diseases such as Alzheimer's disease (AD). Cdk5 phosphorylates tau at AD-specific phospho-epitopes when it associates with p25. p25 is a truncated activator, which is produced from the physiological Cdk5 activator p35 upon exposure to Abeta peptides. We show that neuronal infections with Cdk5 inhibitory peptide (CIP) selectively inhibit p25/Cdk5 activity and suppress the aberrant tau phosphorylation in cortical neurons. Furthermore, Abeta(1-42)-induced apoptosis of these cortical neurons was also reduced by coinfection with CIP. Of particular importance is our finding that CIP did not inhibit endogenous or transfected p35/Cdk5 activity, nor did it inhibit the other cyclin-dependent kinases such as Cdc2, Cdk2, Cdk4 and Cdk6. These results, therefore, provide a strategy to address, and possibly ameliorate, the pathology of neurodegenerative diseases that may be a consequence of aberrant p25 activation of Cdk5, without affecting 'normal' Cdk5 activity.

A Cdk5 inhibitory peptide reduces tau hyperphosphorylation and apoptosis in neurons.

The culture of human embryonic stem cells (hESCs) is limited, both technically and with respect to clinical potential, by the use of mouse embryonic fibroblasts (MEFs) as a feeder layer. The concern over xenogeneic contaminants from the mouse feeder cells may restrict transplantation to humans and the variability in MEFs from batch-to-batch and laboratory-to-laboratory may contribute to some of the variability in experimental results. Finally, use of any feeder layer increases the work load and subsequently limits the large-scale culture of human ES cells. Thus, the development of feeder-free cultures will allow more reproducible culture conditions, facilitate scale-up and potentiate the clinical use of cells differentiated from hESC cultures. In this review, we describe various methods tested to culture cells in the absence of MEF feeder layers and other advances in eliminating xenogeneic products from the culture system.

Toward xeno-free culture of human embryonic stem cells.

Species may compete indirectly by altering the traits of a shared resource. For example, herbivore-induced responses in plants may make plants more resistant or susceptible to additional herbivorous insect species. Herbivore-induced plant responses can significantly affect interspecific competition and herbivore population dynamics. These herbivore-herbivore indirect interactions have been overlooked in aquatic ecosystems where previous studies used the same herbivore species to induce changes and to assess the effects of these changes. We asked whether seaweed grazing by one of two herbivorous, congeneric snail species (Littorina obtusata or Littorina littorea) with different feeding strategies and preferences would affect subsequent feeding preferences of three herbivore species (both snails and the isopod Idotea baltica) and population densities of three herbivore species (both snails and a third periwinkle snail, Lacuna vincta). In addition, we measured phlorotannin concentrations to test the hypothesis that these metabolites function as induced defenses in the Phaeophyceae. Snail herbivory induced cue-specific responses in apical tissues of the seaweed Fucus vesiculosus that affected the three herbivore species similarly. When compared to ungrazed controls, direct grazing by Littorina obtusata reduced seaweed palatability by at least 52% for both snail species and the isopod species. In contrast, direct grazing by L. littorea did not decrease seaweed palatability for any herbivore, indicating herbivore-specific responses. Previous grazing by L. obtusata reduced populations of L. littorea on outplanted seaweeds by 46% but had no effect on L. obtusata populations. Phlorotannins, a potential class of inducible chemicals in brown algae, were not more concentrated in grazed seaweed tissues, suggesting that some other trait was responsible for the induced resistance. Our results indicate that marine herbivores may compete via inducible responses in shared seaweeds. These plant-mediated interactions were asymmetric with a specialist (L. obtusata) competitively superior to a generalist (L. littorea).

Asymmetric competition via induced resistance: specialist herbivores indirectly suppress generalist preference and populations.

The International Stem Cell Initiative characterized 59 human embryonic stem cell lines from 17 laboratories worldwide. Despite diverse genotypes and different techniques used for derivation and maintenance, all lines exhibited similar expression patterns for several markers of human embryonic stem cells. They expressed the glycolipid antigens SSEA3 and SSEA4, the keratan sulfate antigens TRA-1-60, TRA-1-81, GCTM2 and GCT343, and the protein antigens CD9, Thy1 (also known as CD90), tissue-nonspecific alkaline phosphatase and class 1 HLA, as well as the strongly developmentally regulated genes NANOG, POU5F1 (formerly known as OCT4), TDGF1, DNMT3B, GABRB3 and GDF3. Nevertheless, the lines were not identical: differences in expression of several lineage markers were evident, and several imprinted genes showed generally similar allele-specific expression patterns, but some gene-dependent variation was observed. Also, some female lines expressed readily detectable levels of XIST whereas others did not. No significant contamination of the lines with mycoplasma, bacteria or cytopathic viruses was detected.

Characterization of human embryonic stem cell lines by the International Stem Cell Initiative.

The expression and function of several multidrug transporters (including ABCB1 and ABCG2) have been studied in human cancer cells and in mouse and human adult stem cells. However, the expression of ABCG2 in human embryonic stem cells (hESCs) remains unclear. Limited and contradictory results in the literature from two research groups have raised questions regarding its expression and function. In this study, we used quantitative real-time PCR, Northern blots, whole genome RNA sequencing, Western blots, and immunofluorescence microscopy to study ABCG2 expression in hESCs. We found that full-length ABCG2 mRNA transcripts are expressed in undifferentiated hESC lines. However, ABCG2 protein was undetectable even under embryoid body differentiation or cytotoxic drug induction. Moreover, surface ABCG2 protein was coexpressed with the differentiation marker stage-specific embryonic antigen-1 of hESCs, following constant BMP-4 signaling at days 4 and 6. This expression was tightly correlated with the downregulation of two microRNAs (miRNAs) (i.e., hsa-miR-519c and hsa-miR-520h). Transfection of miRNA mimics and inhibitors of these two miRNAs confirmed their direct involvement in the regulation ABCG2 translation. Our findings clarify the controversy regarding the expression of the ABCG2 gene and also provide new insights into translational control of the expression of membrane transporter mRNAs by miRNAs in hESCs.

Regulation and expression of the ATP-binding cassette transporter ABCG2 in human embryonic stem cells.

Regenerative medicine, relying on human embryonic stem cell (hESC) technology, opens promising new avenues for therapy of many severe diseases. However, this approach is restricted by limited production of the desired cells due to the refractory properties of hESC growth in vitro. It is further hindered by insufficient control of cellular stress, growth rates, and heterogeneous cellular states under current culture conditions. In this study, we report a novel cell culture method based on a non-colony type monolayer (NCM) growth. Human ESCs under NCM remain pluripotent as determined by teratoma assays and sustain the potential to differentiate into three germ layers. This NCM culture has been shown to homogenize cellular states, precisely control growth rates, significantly increase cell production, and enhance hESC recovery from cryopreservation without compromising chromosomal integrity. This culture system is simple, robust, scalable, and suitable for high-throughput screening and drug discovery.

Non-colony type monolayer culture of human embryonic stem cells.

&#x5F88;&#x591A;&#x7531;&#x8BF1;&#x5BFC;&#x7684;&#x591A;&#x529F;&#x80FD;&#x5E72;&#x7EC6;&#x80DE;&#x6280;&#x672F;&#x751F;&#x6210;&#x7684;&#x5174;&#x594B;&#x4E0E;&#x5EFA;&#x6A21;&#x4EE5;&#x53CA;&#x6F5C;&#x5728;&#x7684;&#x4E2A;&#x6027;&#x5316;&#x7684;&#x7EC6;&#x80DE;&#x6CBB;&#x7597;&#x7684;&#x75BE;&#x75C5;&#x7684;&#x53EF;&#x80FD;&#x6027;&#x6709;&#x5173;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x8981;&#x8FFD;&#x6C42;&#x8FD9;&#x662F;&#x91CD;&#x8981;&#x7684;&#x662F;&#x8981;&#x4E86;&#x89E3;&#x5305;&#x62EC;&#x5176;&#x56FA;&#x6709;&#x53D8;&#x5F02;&#x6027;&#x591A;&#x80FD;&#x5E72;&#x6B63;&#x5E38;&#x72B6;&#x6001;&#x3002;&#x6211;&#x4EEC;&#x6267;&#x884C;&#x5404;&#x79CD;&#x5206;&#x5B50;&#x5206;&#x6790;&#x5316;&#x9A8C; 21 hESC &#x7EBF;&#x6761;&#x548C; 8 hiPSC &#x884C;&#x6765;&#x751F;&#x6210;&#x5168;&#x9762;&#x7684;&#x591A;&#x80FD;&#x5E72;&#x7EC6;&#x80DE;&#x7684;&#x672A;&#x5206;&#x5316;&#x72B6;&#x6001;&#x7684;&#x5FEB;&#x7167;&#x3002;&#x57FA;&#x56E0;&#x8868;&#x8FBE;&#x6570;&#x636E;&#x7684;&#x5206;&#x6790;&#x663E;&#x793A;&#xFF0C;&#x6839;&#x636E;&#x524D;&#x51E0;&#x6B21;&#x62A5;&#x544A;&#x6CA1;&#x6709;&#x673A;&#x4F53;&#x529F;&#x80FD;&#x7279;&#x5F02;&#x6027;&#x57FA;&#x56E0;&#x8868;&#x8FBE;&#x6A21;&#x5F0F;&#x3002;&#x6211;&#x4EEC;&#x8FDB;&#x4E00;&#x6B65;&#x6BD4;&#x8F83;&#x5206;&#x5316;&#x4F5C;&#x4E3A;&#x80DA;&#x4F53; 2 &#x5A92;&#x4F53;&#x62DF;&#x542F;&#x52A8;&#x5BF9;&#x5355;&#x72EC;&#x7684;&#x5355;&#x5143;&#x683C;&#x7684;&#x547D;&#x8FD0;&#xFF0C;&#x4EE5;&#x53CA; 20 &#x6210;&#x4EBA;&#x7EC4;&#x7EC7;&#x5206;&#x5316;&#x7684;&#x7EC6;&#x80DE;&#x3002;&#x4ECE;&#x8FD9;&#x4E00;&#x5206;&#x6790;&#x6211;&#x4EEC;&#x5DF2;&#x7ECF;&#x4EA7;&#x751F;&#x4E86;&#x57FA;&#x56E0;&#x5217;&#x8868;&#x5B9A;&#x4E49;&#x591A;&#x6F5C;&#x80FD;&#xFF0C;&#x4E3A;&#x591A;&#x80FD;&#x5E72;&#x56FD;&#x5BB6;&#x5EFA;&#x7ACB;&#x4E00;&#x4E2A;&#x57FA;&#x7EBF;&#x3002;&#x6700;&#x540E;&#xFF0C;&#x6211;&#x4EEC;&#x63D0;&#x4F9B;&#x5217;&#x8868;&#x663E;&#x793A;&#x72EC;&#x7ACB;&#x5355;&#x5143;&#x683C;&#x547D;&#x8FD0;&#x7684;&#x62DF;&#x8BAE;&#x7684;&#x504F;&#x89C1;&#x8FD9;&#x4E24;&#x79CD;&#x5206;&#x5316;&#x6761;&#x4EF6;&#x4E30;&#x5BCC;&#x7684;&#x57FA;&#x56E0;&#x3002;

StemCellDB&#xFF1A; &#x4EBA;&#x7C7B;&#x80DA;&#x80CE;&#x5E72;&#x7EC6;&#x80DE;&#x6570;&#x636E;&#x5E93;&#x5728;&#x56FD;&#x5BB6;&#x5065;&#x5EB7;&#x7814;&#x7A76;&#x6240;&#x3002;

Much of the excitement generated by induced pluripotent stem cell technology is concerned with the possibility of disease modeling as well as the potential for personalized cell therapy. However, to pursue this it is important to understand the 'normal' pluripotent state including its inherent variability. We have performed various molecular profiling assays for 21 hESC lines and 8 hiPSC lines to generate a comprehensive snapshot of the undifferentiated state of pluripotent stem cells. Analysis of the gene expression data revealed no iPSC-specific gene expression pattern in accordance with previous reports. We further compared cells, differentiated as embryoid bodies in 2 media proposed to initiate differentiation towards separate cell fates, as well as 20 adult tissues. From this analysis we have generated a gene list which defines pluripotency and establishes a baseline for the pluripotent state. Finally, we provide lists of genes enriched under both differentiation conditions which show the proposed bias toward independent cell fates.

StemCellDB: The Human Pluripotent Stem Cell Database at the National Institutes of Health.

Une grande partie de l&#39;enthousiasme g&eacute;n&eacute;r&eacute; par la technologie des cellules souches pluripotentes induites est pr&eacute;occup&eacute;e par la possibilit&eacute; de maladie de mod&eacute;lisation ainsi que le potentiel de la th&eacute;rapie cellulaire personnalis&eacute;. Toutefois, afin de poursuivre ce, il est important de comprendre l&#39;&eacute;tat de pluripotentes &laquo; normal &raquo;, y compris sa variabilit&eacute; inh&eacute;rente. Nous avons effectu&eacute; plusieurs essais mol&eacute;culaires de profilage pour 21 lign&eacute;es de CSEh et 8 lignes de hiPSC pour g&eacute;n&eacute;rer un aper&ccedil;u complet de l&#39;&eacute;tat indiff&eacute;renci&eacute; de cellules souches pluripotentes. Analyse les donn&eacute;es d&#39;expression g&eacute;nique a r&eacute;v&eacute;l&eacute; aucun mod&egrave;le d&#39;expression sp&eacute;cifiques iPSC g&egrave;ne conform&eacute;ment aux pr&eacute;c&eacute;dents rapports. Plus loin, nous avons compar&eacute; les cellules diff&eacute;renci&eacute;es comme corps embryo&iuml;des dans 2 m&eacute;dias propos&eacute;s de lancer la diff&eacute;renciation vers des destins cellulaires distincts, ainsi que 20 tissus adultes. De cette analyse, nous avons g&eacute;n&eacute;r&eacute; une liste de g&egrave;ne qui d&eacute;finit la pluripotence et &eacute;tablit une ligne de base pour l&#39;&eacute;tat pluripotent. Enfin, nous fournissons des listes de g&egrave;nes enrichis dans les deux conditions de diff&eacute;renciation qui montrent le biais propos&eacute; vers les destins de cellules ind&eacute;pendantes.

StemCellDB : La base de donn&eacute;es de cellules souches pluripotentes humaines au niveau National Institutes of Health.

Viel von der Aufregung von induzierten pluripotenten Stammzellen-Technologie befasst sich mit der M&ouml;glichkeit der Modellierung sowie das Potenzial f&uuml;r personalisierte Zelltherapie-Krankheit. Um dies zu verfolgen ist es jedoch wichtig zu verstehen, den &#39;normalen&#39; pluripotenten Zustand einschlie&szlig;lich ihrer inh&auml;renten Variabilit&auml;t. Wir haben verschiedene Molekulare Profilerstellungs-Assays f&uuml;r 21 hESC-Linien und 8 HiPSC Zeilen generieren einen umfassenden Snapshot des undifferenzierten pluripotenten Stammzellen durchgef&uuml;hrt. Analyse der Gen-Ausdruck-Daten ergab kein iPSC-spezifische Gen-Ausdruck-Muster nach fr&uuml;heren Berichten. Wir verglichen weitere Zellen, differenzierte als Embryoid K&ouml;rper in 2 Medien vorgeschlagene Differenzierung gegen&uuml;ber separate Zelle Schicksale sowie 20 Erwachsene Gewebe zu initiieren. Aus dieser Analyse haben wir eine gen-Liste erzeugt, die Pluripotenz definiert und etabliert eine Baseline f&uuml;r den pluripotenten Zustand. Schlie&szlig;lich bieten wir Listen von Genen, die unter beiden Differenzierung-Bedingungen, die die vorgeschlagenen in Richtung unabh&auml;ngiger Zelle Schicksale Einseitigkeit bereichert.

StemCellDB: Die menschliche Pluripotent Stem Cell Datenbank an den National Institutes of Health.

Molto l&#39;entusiasmo suscitato dalla tecnologia delle cellule staminali pluripotenti indotte &egrave; interessato la possibilit&agrave; di malattia modellazione cos&igrave; come il potenziale per la terapia cellulare personalizzato. Tuttavia, per perseguire questo &egrave; importante comprendere lo stato &#39;normale&#39; pluripotenti compresa sua variabilit&agrave; intrinseca. Abbiamo eseguito vari saggi di analisi molecolare per 21 linee di hESC e 8 hiPSC per generare uno snapshot completo dello stato indifferenziato di cellule staminali pluripotenti. Analisi dei dati di espressione genica ha rivelato nessun modello di espressione iPSC specifico gene conformemente a precedenti relazioni. Abbiamo confrontato ulteriormente le cellule, differenziate come corpi embryoid in 2 media proposto di avviare la differenziazione verso destini cella separata, come pure i tessuti adulti 20. Da questa analisi abbiamo abbiamo generato un elenco di gene che definisce la pluripotenza e stabilisce una linea di base per lo stato pluripotenti. Infine, forniamo liste di geni arricchiti sotto entrambe le condizioni di differenziazione che mostrano il bias proposto verso destini cellula indipendente.

StemCellDB: L&#39;umano cellule staminali pluripotenti Database presso i National Institutes of Health.

&#x8A98;&#x5C0E;&#x591A;&#x80FD;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#x6280;&#x8853;&#x306B;&#x3088;&#x3063;&#x3066;&#x751F;&#x6210;&#x3055;&#x308C;&#x305F;&#x8208;&#x596E;&#x306E;&#x591A;&#x304F;&#x306F;&#x30D1;&#x30FC;&#x30BD;&#x30CA;&#x30E9;&#x30A4;&#x30BA;&#x3055;&#x308C;&#x305F;&#x7D30;&#x80DE;&#x7642;&#x6CD5;&#x306E;&#x53EF;&#x80FD;&#x6027;&#x3060;&#x3051;&#x3067;&#x306A;&#x304F;&#x3001;&#x30E2;&#x30C7;&#x30EA;&#x30F3;&#x30B0;&#x75BE;&#x60A3;&#x306E;&#x53EF;&#x80FD;&#x6027;&#x3092;&#x61F8;&#x5FF5;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x305F;&#x3060;&#x3057;&#x3001;&#x3053;&#x308C;&#x3092;&#x8FFD;&#x6C42;&#x3059;&#x308B;&#x305D;&#x306E;&#x56FA;&#x6709;&#x306E;&#x5909;&#x52D5;&#x3092;&#x542B;&#x3080; &#39;normal&#39; &#x591A;&#x80FD;&#x6027;&#x306E;&#x72B6;&#x614B;&#x3092;&#x7406;&#x89E3;&#x3059;&#x308B;&#x3053;&#x3068;&#x304C;&#x91CD;&#x8981;&#x3067;&#x3059;&#x3002;&#x69D8;&#x3005; &#x306A;&#x5206;&#x5B50;&#x30A2;&#x30C3;&#x30BB;&#x30A4; 21 hESC &#x7DDA;&#x3068;&#x591A;&#x80FD;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#x306E;&#x672A;&#x5206;&#x5316;&#x72B6;&#x614B;&#x306E;&#x5305;&#x62EC;&#x7684;&#x306A;&#x30B9;&#x30CA;&#x30C3;&#x30D7;&#x30B7;&#x30E7;&#x30C3;&#x30C8;&#x3092;&#x751F;&#x6210;&#x3059;&#x308B; 8 &#x304B;&#x3089;&#x4F5C;&#x88FD;&#x3057;&#x305F;&#x30E9;&#x30A4;&#x30F3;&#x3092;&#x884C;&#x3063;&#x305F;&#x3002;&#x907A;&#x4F1D;&#x5B50;&#x767A;&#x73FE;&#x30C7;&#x30FC;&#x30BF;&#x306E;&#x89E3;&#x6790;&#x306F;&#x3001;&#x4EE5;&#x524D;&#x306E;&#x30EC;&#x30DD;&#x30FC;&#x30C8;&#x306B;&#x5F93;&#x3063;&#x3066; iPSC &#x7279;&#x7570;&#x7684;&#x907A;&#x4F1D;&#x5B50;&#x8868;&#x73FE;&#x30D1;&#x30BF;&#x30FC;&#x30F3;&#x3092;&#x660E;&#x3089;&#x304B;&#x306B;&#x3002;&#x6211;&#x3005; &#x306F;&#x3055;&#x3089;&#x306B;&#x500B;&#x5225;&#x7D30;&#x80DE;&#x904B;&#x547D;&#x3068;&#x3057;&#x3066; 20 &#x6210;&#x4F53;&#x7D44;&#x7E54;&#x3078;&#x306E;&#x5206;&#x5316;&#x3092;&#x958B;&#x59CB;&#x3059;&#x308B;&#x63D0;&#x6848; 2 &#x306E;&#x30E1;&#x30C7;&#x30A3;&#x30A2;&#x306E;&#x80DA;&#x69D8;&#x4F53;&#x3068;&#x3057;&#x3066;&#x5206;&#x5316;&#x3057;&#x305F;&#x7D30;&#x80DE;&#x3092;&#x6BD4;&#x8F03;&#x3057;&#x307E;&#x3057;&#x305F;&#x3002;&#x3053;&#x306E;&#x5206;&#x6790;&#x304B;&#x3089;&#x306F;&#x591A;&#x80FD;&#x6027;&#x3092;&#x5B9A;&#x7FA9;&#x3057;&#x3001;&#x591A;&#x80FD;&#x6027;&#x306E;&#x72B6;&#x614B;&#x306E;&#x305F;&#x3081;&#x306E;&#x30D9;&#x30FC;&#x30B9;&#x30E9;&#x30A4;&#x30F3;&#x3092;&#x78BA;&#x7ACB;&#x3059;&#x308B;&#x907A;&#x4F1D;&#x5B50;&#x30EA;&#x30B9;&#x30C8;&#x3092;&#x751F;&#x6210;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x6700;&#x5F8C;&#x306B;&#x3001;&#x63D0;&#x6848;&#x3055;&#x308C;&#x305F;&#x72EC;&#x7ACB;&#x3057;&#x305F;&#x7D30;&#x80DE;&#x904B;&#x547D;&#x30D0;&#x30A4;&#x30A2;&#x30B9;&#x3092;&#x793A;&#x3059;&#x4E21;&#x65B9;&#x306E;&#x5206;&#x5316;&#x306E;&#x6761;&#x4EF6;&#x306E;&#x4E0B;&#x3067;&#x6FC3;&#x7E2E;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x30EA;&#x30B9;&#x30C8;&#x3092;&#x63D0;&#x4F9B;&#x3057;&#x307E;&#x3059;&#x3002;

StemCellDB&#xFF1A; &#x56FD;&#x7ACB;&#x885B;&#x751F;&#x7814;&#x7A76;&#x6240;&#x3067;&#x306E;&#x3072;&#x3068;&#x591A;&#x80FD;&#x6027;&#x5E79;&#x7D30;&#x80DE;&#x30C7;&#x30FC;&#x30BF;&#x30D9;&#x30FC;&#x30B9;&#x3002;

&#xC720;&#xB3C4; &#xB9CC;&#xB2A5; &#xC904;&#xAE30; &#xC138;&#xD3EC; &#xAE30;&#xC220;&#xC5D0; &#xC758;&#xD574; &#xC0DD;&#xC131; &#xB41C; &#xD765;&#xBD84;&#xC758; &#xC5F4;&#xAE30;&#xAC00; &#xB9CE;&#xC740; &#xC9C8;&#xBCD1; &#xBFD0;&#xB9CC; &#xC544;&#xB2C8;&#xB77C;, &#xB9DE;&#xCDA4;&#xD615;&#xB41C; &#xC138;&#xD3EC; &#xCE58;&#xB8CC;&#xC5D0; &#xB300; &#xD55C; &#xAC00;&#xB2A5;&#xC131; &#xBAA8;&#xB378;&#xB9C1;&#xC758; &#xAC00;&#xB2A5;&#xC131;&#xACFC; &#xD568;&#xAED8; &#xC6B0;&#xB824; &#xD558;&#xACE0;&#xC788;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;, &#xC774;&#xAC83;&#xC744; &#xCD94;&#xAD6C; &#xD558;&#xB294; &#xADF8;&#xAC83;&#xC740; &#xADF8;&#xAC83;&#xC758; &#xB0B4;&#xC7AC; &#xBCC0;&#xB3D9;&#xC131;&#xC744; &#xD3EC;&#xD568; &#xD55C; &#39;&#xC815;&#xC0C1;&#39; pluripotent &#xC0C1;&#xD0DC;&#xB97C; &#xC774;&#xD574; &#xD558;&#xB294; &#xAC83;&#xC774; &#xC911;&#xC694;&#xC785;&#xB2C8;&#xB2E4;. &#xC6B0;&#xB9AC; 21 hESC &#xB77C;&#xC778;&#xACFC; pluripotent &#xC904;&#xAE30; &#xC138;&#xD3EC;&#xC758; undifferentiated &#xC0C1;&#xD0DC;&#xC758; &#xC885;&#xD569;&#xC801;&#xC778; &#xC2A4;&#xB0C5;&#xC20F;&#xC744; &#xC0DD;&#xC131; 8 hiPSC &#xB77C;&#xC778;&#xC5D0; &#xB300; &#xD55C; &#xB2E4;&#xC591; &#xD55C; &#xBD84;&#xC790; &#xD504;&#xB85C; &#xD30C;&#xC77C;&#xB9C1; &#xBD84;&#xC11D; &#xC2E4;&#xD5D8;&#xC744; &#xC218;&#xD589; &#xD588;&#xC2B5;&#xB2C8;&#xB2E4;. &#xC720;&#xC804;&#xC790; &#xD45C;&#xD604; &#xB370;&#xC774;&#xD130; &#xBD84;&#xC11D; iPSC &#xD2B9;&#xC815; &#xC720;&#xC804;&#xC790; &#xC2DD; &#xD328;&#xD134;&#xC744; &#xB530;&#xB77C; &#xC774;&#xC804; &#xBCF4;&#xACE0;&#xC11C;&#xB97C; &#xACF5;&#xAC1C; &#xD588;&#xB2E4;. &#xC6B0;&#xB9AC;&#xB294; &#xB354; &#xC140;, 2 &#xBBF8;&#xB514;&#xC5B4; 20 &#xC131;&#xC778; &#xC870;&#xC9C1;&#xC73C;&#xB85C; &#xBCC4;&#xB3C4; &#xC140; &#xC6B4;&#xBA85;&#xC744; &#xD5A5;&#xD574; &#xCC28;&#xBCC4;&#xD654; &#xC2DC;&#xC791; &#xC81C;&#xC548; embryoid &#xC2DC;&#xCCB4;&#xB85C; &#xCC28;&#xBCC4;&#xD654; &#xB41C; &#xBE44;&#xAD50;. &#xC774; &#xBD84;&#xC11D;&#xC5D0;&#xC11C; &#xC6B0;&#xB9AC; &#xC815;&#xC758; pluripotency pluripotent &#xC0C1;&#xD0DC;&#xC5D0; &#xB300; &#xD55C; &#xCD08;&#xAE30; &#xACC4;&#xD68D;&#xC744; &#xC124;&#xC815; &#xD558;&#xB294; &#xC720;&#xC804;&#xC790; &#xBAA9;&#xB85D;&#xC744; &#xC0DD;&#xC131;. &#xB9C8;&#xC9C0;&#xB9C9;&#xC73C;&#xB85C;, &#xC6B0;&#xB9AC;&#xB294; &#xB3C5;&#xB9BD;&#xC801;&#xC778; &#xC138;&#xD3EC; &#xC6B4;&#xBA85; &#xD5A5;&#xD574; &#xC81C;&#xC548; &#xB41C; &#xBC14;&#xC774;&#xC5B4;&#xC2A4;&#xB97C; &#xBCF4;&#xC5EC; &#xB450; &#xBD84;&#xD654; &#xC870;&#xAC74; &#xD558;&#xC5D0;&#xC11C; &#xB18D;&#xCD95; &#xD558;&#xB294; &#xC720;&#xC804;&#xC790;&#xC758; &#xBAA9;&#xB85D;&#xC744; &#xC81C;&#xACF5; &#xD569;&#xB2C8;&#xB2E4;.

StemCellDB: &#xC778;&#xAC04;&#xC758; Pluripotent &#xC904;&#xAE30; &#xC138;&#xD3EC; &#xB370;&#xC774;&#xD130;&#xBCA0;&#xC774;&#xC2A4; &#xAC74;&#xAC15;&#xC758; &#xAD6D;&#xB9BD; &#xC5F0;&#xAD6C;&#xC18C;&#xC5D0;&#xC11C;.

Gran parte de la emoci&oacute;n generada por la tecnolog&iacute;a de c&eacute;lulas madre pluripotentes inducidas se refiere a la posibilidad de enfermedad modelado as&iacute; como el potencial de la terapia celular personalizado. Sin embargo, para dedicarse a esto es importante comprender el estado pluripotente &#39;normal&#39;, incluyendo su variabilidad inherente. Hemos realizado varios ensayos perfiles moleculares para 21 hESCs y 8 hiPSC l&iacute;neas al generar una instant&aacute;nea completa del estado indiferenciado de c&eacute;lulas madre pluripotentes. El an&aacute;lisis de los datos de expresi&oacute;n gen&eacute;tica no revel&oacute; ning&uacute;n patr&oacute;n de expresi&oacute;n de genes espec&iacute;ficos de iPSC seg&uacute;n informes anteriores. Comparamos m&aacute;s c&eacute;lulas diferenciadas como cuerpos embryoid en 2 medios propuestos iniciar la diferenciaci&oacute;n hacia destinos separados de la c&eacute;lula, as&iacute; como 20 tejidos adultos. De este an&aacute;lisis hemos generado una lista del gen que define la pluripotencia y establece un punto de partida para el estado pluripotente. Por &uacute;ltimo, proporcionamos listas de genes enriquecidos en ambas condiciones de diferenciaci&oacute;n que demuestran el sesgo propuesto hacia destinos de c&eacute;lulas independientes.

StemCellDB: El humano Pluripotent Stem Cell base en los institutos nacionales de salud.

Many studies have compared the genetic and epigenetic profiles of human induced pluripotent stem cells (hiPSCs) to human embryonic stem cells (hESCs) and yet the picture remains unclear. To address this, we derived a population of neural precursor cells (NPCs) from the H1 (WA01) hESC line and generated isogenic iPSC lines by reprogramming. The gene expression and methylation profile of three lines were compared to the parental line and intermediate NPC population. We found no gene probe with expression that differed significantly between hESC and iPSC samples under undifferentiated or differentiated conditions. Analysis of the global methylation pattern also showed no significant difference between the two PSC populations. Both undifferentiated populations were distinctly different from the intermediate NPC population in both gene expression and methylation profiles. One point to note is that H1 is a male line and so extrapolation to female lines should be cautioned. However, these data confirm our previous findings that there are no significant differences between hESCs and hiPSCs at the gene expression or methylation level.

Comparison of the molecular profiles of human embryonic and induced pluripotent stem cells of isogenic origin.

Human pluripotent stem cells (hPSCs) have two potentially attractive applications: cell replacement-based therapies and drug discovery. Both require the efficient generation of large quantities of clinical-grade stem cells that are free from harmful genomic alterations. The currently employed colony-type culture methods often result in low cell yields, unavoidably heterogeneous cell populations, and substantial chromosomal abnormalities. Here, we shed light on the structural relationship between hPSC colonies/embryoid bodies and early-stage embryos in order to optimize current culture methods based on the insights from developmental biology. We further highlight core signaling pathways that underlie multiple epithelial-to-mesenchymal transitions (EMTs), cellular heterogeneity, and chromosomal instability in hPSCs. We also analyze emerging methods such as non-colony type monolayer (NCM) and suspension culture, which provide alternative growth models for hPSC expansion and differentiation. Furthermore, based on the influence of cell-cell interactions and signaling pathways, we propose concepts, strategies, and solutions for production of clinical-grade hPSCs, stem cell precursors, and miniorganoids, which are pivotal steps needed for future clinical applications.

Developmental insights from early mammalian embryos and core signaling pathways that influence human pluripotent cell growth and differentiation.

The ability to differentiate induced pluripotent stem cells (iPSCs) into committed skeletal progenitors could allow for an unlimited autologous supply of such cells for therapeutic uses; therefore, we attempted to create novel bone-forming cells from human iPSCs using lines from two distinct tissue sources and methods of differentiation that we previously devised for osteogenic differentiation of human embryonic stem cells, and as suggested by other publications. The resulting cells were assayed using in vitro methods, and the results were compared with those obtained from in vivo transplantation assays. Our results show that true bone was formed in vivo by derivatives of several iPSC lines, but that the successful cell lines and differentiation methodologies were not predicted by the results of the in vitro assays. In addition, bone was formed equally well from iPSCs originating from skin or bone marrow stromal cells (also known as bone marrow-derived mesenchymal stem cells), suggesting that the iPSCs did not retain a "memory" of their previous life. Furthermore, one of the iPSC-derived cell lines formed verifiable cartilage in vivo, which likewise was not predicted by in vitro assays.

Rebecca S. Hamilton

NIH Stem Cell Unit,
National Institute of Neurological Disorders and Stroke,
NIH Stem Cell Unit, National Institute of Neurological Disorders and Stroke

National Institutes of Health

Alternative Cultures for Human Pluripotent Stem Cell Production, Maintenance, and Genetic Analysis

Rebecca S. Hamilton

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}NIH Stem Cell Unit,National Institute of Neurological Disorders and Stroke,NIH Stem Cell Unit, National Institute of Neurological Disorders and Stroke

National Institutes of Health

Alternative Cultures for Human Pluripotent Stem Cell Production, Maintenance, and Genetic Analysis

NIH Stem Cell Unit,
National Institute of Neurological Disorders and Stroke,
NIH Stem Cell Unit, National Institute of Neurological Disorders and Stroke