Name: transfection, selection, and colony-picking of human induced pluripotent stem cells talen-targeted with a gfp gene into the aavs1 safe harbor
Uploaded: 2015-02-01T14:00:00
Description: Watch this Scientific Journal Video about Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor at JoVE.com

This research was supported by the NIH Common Fund and Intramural Research Program of the National Institute of Arthritis, Musculoskeletal, and Skin Diseases.

1. Preparazione della matrice di membrana basale e rivestimento dei Plasticware <ol><li> Posizionare il congelato membrana basale matrice magazzino da -20 ° C su ghiaccio e scongelare una notte a 4 ° C. </li><li> Dopo lo scongelamento, pipetta 2 mg aliquote di matrice membrana basale in tubi Eppendorf pre-refrigerati. Conservare questi a -20 ° C fino al momento dell&#39;uso. </li><li> Per preparare piatti matrice rivestite membrana basale, scongelare un&#39;aliquota sul ghiaccio fino a quando l&#39;ultimo pezzo di ...

<ol>
	<li>Takahashi, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+pluripotent+stem+cells+from+adult+human+fibroblasts+by+defined+factors.">Induction of pluripotent stem cells from adult human fibroblasts by defined factors.</a> Cell. 131 (5), 861-872 (2007).</li><li>Hentze, H., Graichen, R., Colman, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+therapy+and+the+safety+of+embryonic+stem+cell-derived+grafts.">Cell therapy and the safety of embryonic stem cell-derived grafts.</a> Trends Biotechnol. 25 (1), 24-32 (2007).</li><li>Smith, J. 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P., Thomson, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Homologous+recombination+in+human+embryonic+stem+cells.">Homologous recombination in human embryonic stem cells.</a> Nat Biotechnol. 21 (3), 319-321 (2003).</li><li>Urbach, A., Schuldiner, M., Benvenisty, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+for+Lesch-Nyhan+disease+by+gene+targeting+in+human+embryonic+stem+cells.">Modeling for Lesch-Nyhan disease by gene targeting in human embryonic stem cells.</a> Stem Cells. 22 (4), 635-641 (2004).</li><li>Hockemeyer, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+targeting+of+expressed+and+silent+genes+in+human+ESCs+and+iPSCs+using+zinc-finger+nucleases.">Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases.</a> Nature biotechnology. 27 (9), 851-857 (2009).</li><li>Zou, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+targeting+of+a+disease-related+gene+in+human+induced+pluripotent+stem+and+embryonic+stem+cells.">Gene targeting of a disease-related gene in human induced pluripotent stem and embryonic stem cells.</a> Cell stem cell. 5 (1), 97-110 (2009).</li><li>Hockemeyer, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+engineering+of+human+pluripotent+cells+using+TALE+nucleases.">Genetic engineering of human pluripotent cells using TALE nucleases.</a> Nature. 29 (8), 731-734 (2011).</li><li>Sanjana, N. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+transcription+activator-like+effector+toolbox+for+genome+engineering.">A transcription activator-like effector toolbox for genome engineering.</a> Nature protocols. 7 (1), 171-192 (2012).</li><li>Mali, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RNA-guided+human+genome+engineering+via+Cas9.">RNA-guided human genome engineering via Cas9.</a> Science. 339 (6121), 823-826 (2013).</li><li>Ding, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enhanced+efficiency+of+human+pluripotent+stem+cell+genome+editing+through+replacing+TALENs+with+CRISPRs.">Enhanced efficiency of human pluripotent stem cell genome editing through replacing TALENs with CRISPRs.</a> Cell stem cell. 12 (4), 393-394 (2013).</li><li>Luo, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+Enhanced+Green+Fluorescent+Protein+Expression+After+Differentiation+and+Transplantation+of+Reporter+Human.">Stable Enhanced Green Fluorescent Protein Expression After Differentiation and Transplantation of Reporter Human.</a> Induced Pluripotent Stem Cells Generated by AAVS1 Transcription Activator-Like Effector Nucleases. Stem Cells Transl Med. 3 (7), 821-835 (2014).</li><li>Zou, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxidase-deficient+neutrophils+from+X-linked+chronic+granulomatous+disease+iPS+cells:+functional+correction+by+zinc+finger+nuclease-mediated+safe+harbor+targeting.">Oxidase-deficient neutrophils from X-linked chronic granulomatous disease iPS cells: functional correction by zinc finger nuclease-mediated safe harbor targeting.</a> Blood. 117 (21), 5561-5572 (2011).</li><li>Luo, Y., Rao, M., Zou, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+GFP+Reporter+Human+Induced+Pluripotent+Stem+Cells+Using+AAVS1+Safe+Harbor+Transcription+Activator-Like+Effector+Nuclease.">Generation of GFP Reporter Human Induced Pluripotent Stem Cells Using AAVS1 Safe Harbor Transcription Activator-Like Effector Nuclease.</a> Curr Protoc Stem Cell Biol. 29, 5A.7.1-5A.7.18 (2014).</li><li>Ran, F. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genome+engineering+using+the+CRISPR-Cas9+system.">Genome engineering using the CRISPR-Cas9 system.</a> Nat Protoc. 8 (11), 2281-2308 (2013).</li><li>Beers, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Passaging+and+colony+expansion+of+human+pluripotent+stem+cells+by+enzyme-free+dissociation+in+chemically+defined+culture+conditions.">Passaging and colony expansion of human pluripotent stem cells by enzyme-free dissociation in chemically defined culture conditions.</a> Nat Protoc. 7, 2029-2040 (2012).</li></ol>

Le fasi più critiche per il successo generazione di AAVS1 esente mirati iPSCs umani sono: (1) efficiente di Talen e donatori plasmidi in iPSCs by trasfezione; (2) ottimizzare dissociazione di iPSCs nelle singole cellule prima trasfezione e placcatura densità dopo trasfezione; (3) ottimizzare dose ed il tempo di farmaco-selezione basata sulla crescita della linea COPSI; (4) dissezionare con cura e raccolta colonie mirate e trasferimento alla nuova piastra / bene. Rispetto ai metodi simili impiegati per la carta di Hock...

La possibilità di riprogrammare le cellule somatiche umane in cellule staminali pluripotenti indotte cellule simil-staminali embrionali (iPSCs) fu scoperto da Takahashi et al., Nel 2007 1. Fibroblasti dermici umani trasdotte con retrovirus che esprime quattro fattori di trascrizione (Il Yamanaka così soprannominato Fattori Oct3 / 4, Sox2, c-Myc e Klf4) hanno mostrato di essere molto simili alle cellule staminali embrionali umane (hESC) basati sulla morfologia, la proliferazione, l&#39;espressione genica, e lo stato epigenetico; cruciale, iPSCs sono anche in grado di differenziarsi in cellule di tutti e tre i foglietti embrionali 1. Il potenziale proliferativo e la differenziazione della capacità di iPSCs li rende strumenti molto interessanti; riprogrammando cellule di pazienti affetti da malattie specifiche, iPSCs possono essere utilizzati sia come sistemi modello in vitro di malattia e come potenziali terapie. Per quest&#39;ultimo scopo, diverse questioni devono essere affrontate prima che il pieno potenziale di iPSCsin un ambiente clinico può essere realizzato; il potenziale oncogeno di hESC coltivate in vitro e iPSCs, l&#39;uso di derivati ​​xenogeniche durante la riprogrammazione e la manutenzione delle cellule, e la necessità di tenere traccia di cellule trapiantate in vivo sono tutti ostacoli cruciali per l&#39;applicazione clinica delle cellule staminali pluripotenti (Inviato da Hentze et al. 2). Una soluzione ideale per la necessità di monitoraggio cellule differenziate post-trapianto comporterebbe un marcatore visivamente rilevabile che resiste tacere e variegatura indipendentemente dall&#39;applicazione. Espressione robusta e sostenuta di transgeni integrati è più facilmente realizzabile quando il DNA esogeno viene introdotto in porto sicuro loci; cioè, i siti genomici che consentono sufficienti trascrizione di un vettore integrato mentre allo stesso tempo mitigare perturbazioni di espressione in geni vicini 3. Uno di questi siti che è stato molto ben caratterizzati dalla sua scoperta è il virus integr adeno-associatosito zione 1 (AAVS1), nel primo introne della proteina fosfatasi 1 subunità 12C regolamentazione (PPP1R12C) gene. Questo locus è stato dimostrato non solo di permettere sostenuta e robusta espressione dei transgeni integrati attraverso il tempo prolungato nella cultura e nella differenziazione vitro 3, ma anche per proteggere i geni di perturbazione trascrizionale 4 circostante; entrambe le funzioni si pensa sia dovuto alla presenza di elementi endogeni isolante cromatina che fiancheggiano il sito AAVS1 5. Advances in strumenti di ingegneria del genoma sul proprio nell&#39;ultimo decennio hanno notevolmente facilitato la facilità e l&#39;efficienza con cui le manipolazioni genetiche in qualsiasi tipo di cellula possono essere raggiunti. Mentre i primi esperimenti riusciti invocati estremamente bassi livelli di endogena ricombinazione omologa (HR) con un donatore introdotte per ottenere gene targeting in CES 6,7, l&#39;uso di nucleasi sito-specifiche, quali nucleasi dita di zinco (ZFNs), che signi, significativamente indurre ricombinazione omologa attraverso la generazione di un doppio filamento rottura del DNA ha notevolmente aumentato l&#39;efficienza di tali esperimenti 8,9. La riproposizione di entrambi trascrizione attivatore come effettori (racconti) di pianta patogeno xanthomonas generi e le procariote cluster brevi ripetizioni palindromiche regolarmente intervallati (CRISPR) Sistema / Cas9 in efficienti nucleasi progettista site specific ha fatto gene targeting nelle cellule staminali pluripotenti un accessibile e metodologia possibile 10-13. Un recente ha descritto un metodo efficace per l&#39;integrazione stabile di una cassetta reporter fluorescente verde nel AAVS1 porto sicuro locus in iPSCs umani con nucleasi Tale (Talens) 14. Questi iPSCs mirati hanno mantenuto la loro fluorescenza, anche dopo la differenziazione diretta a cardiomiociti e il trapianto in un modello murino di infarto del miocardio (MI), che fornisce una forte evidenza per l&#39;utilità di talestabilmente pluripotenti fluorescente cellule staminali 14. Per ottenere colonie mirati, un metodo gene-trap è stato usato cui un splicing-accettore (SA), 2A sequenza peptidica auto-scissione colloca la puromicina N-acetil-transferasi (PAC) gene sotto il controllo del promotore PPP1R12C endogena; quindi, solo iPSCs che hanno incorporato il donatore DNA a livello del locus AAVS1 esprimono PAC, rendendoli selezionabili in base puromicina-resistenza; (Figura 1, 15). Dettagli Questo protocollo le procedure di generare AAVS1-GFP iPSCs segnalato nel recente documento 14, tra cui il processo di trasfettando iPSCs con Talens e 9,8 kb donatori di integrare un frammento di 4.2kb DNA nel locus AAVS1 porto sicuro, selezionando iPSCs basati su puromicina-resistenza, e le colonie di prelievo per l&#39;espansione clonale. Le tecniche qui descritte possono essere applicate a vari esperimenti di ingegneria genoma.

<table border="0" cellpadding="0" cellspacing="0" width="1437">
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		<tr height="21">
			<td height="21" style="height:21px;width:588px;">NAME OF MATERIAL/EQUIPMENT</td>
			<td style="width:123px;">COMPANY</td>
			<td style="width:123px;">CATALOG #</td>
			<td style="width:603px;">COMMENTS/DESCRIPTION</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Matrigel Growth Factor Reduced (GFR) Basement Membrane Matrix, *LDEV-Free, 10mL</td>
			<td>Corning</td>
			<td>354230</td>
			<td>Store at -20&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DMEM/F-12</td>
			<td>Life Technologies</td>
			<td>11320-033</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Costar 6 Well Clear TC-Treated Multiple Well Plates</td>
			<td>Corning</td>
			<td>3506</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Essential 8 Medium</td>
			<td>Life Technologies</td>
			<td>A1517001</td>
			<td>Store basal medium at 4&#176;C. Store supplement at -20&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Y-27632 dihydrochloride</td>
			<td>Tocris</td>
			<td>1254</td>
			<td>Store at room temp. Once dissolved in H2O, store at -20&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sodium Chloride</td>
			<td>Sigma</td>
			<td>S5886-500G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">UltraPure 0.5M EDTA, pH 8.0</td>
			<td>Life Technologies</td>
			<td>15575-020</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DPBS, no calcium, no magnesium</td>
			<td>Life Technologies</td>
			<td>14190-250</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">100mm TC-Treated Culture Dish</td>
			<td>Corning</td>
			<td>430167</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DR4 MEF 2M IRR - Academic</td>
			<td>GlobalStem</td>
			<td>GSC-6204G</td>
			<td>Store in liquid Nitrogen.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">DMEM, high glucose, pyruvate</td>
			<td>Life Technologies</td>
			<td>11995-040</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Defined Fetal Bovine Serum, US Origin</td>
			<td>HyClone</td>
			<td>SH30070.03</td>
			<td>Store at -20&#176;C. Thaw at 4&#176;C overnight and aliquot. Store aliquots at -20&#176;C until needed.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MEM Non-Essential Amino Acids Solution (100X)</td>
			<td>Life Technologies</td>
			<td>11140-050</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4D-Nucleofector Core unit&#160;</td>
			<td>Lonza</td>
			<td>AAF-1001B</td>
			<td>part of the electroporation system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">4D-Nucleofector X unit&#160;</td>
			<td>Lonza</td>
			<td>AAF-1001X</td>
			<td>part of the electroporation system</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">P3 Primary Cell 4D-Nucleofector X Kit L (24 RCT)</td>
			<td>Lonza</td>
			<td>V4XP-3024</td>
			<td>Upon arrival, remove Primary Cell Solution and supplement and store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">StemPro Accutase Cell Dissociation Reagent</td>
			<td>Life Technologies</td>
			<td>A1110501</td>
			<td>Store at -20&#176;C. Thaw overnight at 4&#176;C and warm an aliquot in a 37&#176;C water bath before use.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">NutriStem XF/FF Culture Medium</td>
			<td>Stemgent</td>
			<td>01-0005</td>
			<td>Store at -20&#176;C. Thaw overnight at 4&#176;C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">AAVS1 TALENs (pZT-AAVS1-L1 and pZT-AAVS1-R1)</td>
			<td>Addgene</td>
			<td>52637 and 52638</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">AAVS1-CAG-EGFP Homologous Recombination donor</td>
			<td>Addgene</td>
			<td>22212</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Puromycin Dihydrochloride</td>
			<td>Life Technologies</td>
			<td>A11138-03</td>
			<td>Store at -20&#176;C. Prepare working aliquots of 1 mg/mL in ddH2O.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Disposable Borosilicate Glass Pasteur Pipets</td>
			<td>Fisher Scientific</td>
			<td>13-678-20A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sorvall Legend XTR (Refrigerated), 120V 60Hz</td>
			<td>Thermo Scientific</td>
			<td>75-004-521</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TX-750 4 &#215; 750mL Swinging Bucket Rotor</td>
			<td>Thermo Scientific</td>
			<td>75003607</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Trypan Blue Solution, 0.4%</td>
			<td>Life Technologies</td>
			<td>15250-061</td>
			<td></td>
		</tr>
	</tbody>
</table>

transfection, selection, and colony-picking of human induced pluripotent stem cells talen-targeted with a gfp gene into the aavs1 safe harbor

Targeted transgene addition can provide persistent gene expression while circumventing the gene silencing and insertional mutagenesis caused by viral vector mediated random integration. This protocol describes a universal and efficient transgene targeted addition platform in human iPSCs based on utilization of validated open-source TALENs and a gene-trap-like donor to deliver transgenes into a safe harbor locus. Importantly, effective gene editing is rate-limited by the delivery efficiency of gene editing vectors. Therefore, this protocol first focuses on preparation of iPSCs for transfection to achieve high nuclear delivery efficiency. When iPSCs are dissociated into single cells using a gentle-cell dissociation reagent and transfected using an optimized program, >50% cells can be induced to take up the large gene editing vectors. Because the AAVS1 locus is located in the intron of an active gene (PPP1R12C), a splicing acceptor (SA)-linked puromycin resistant gene (PAC) was used to select targeted iPSCs while excluding random integration-only and untransfected cells. This strategy greatly increases the chance of obtaining targeted clones, and can be used in other active gene targeting experiments as well. Two weeks after puromycin selection at the dose adjusted for the specific iPSC line, clones are ready to be picked by manual dissection of large, isolated colonies into smaller pieces that are transferred to fresh medium in a smaller well for further expansion and genetic and functional screening. One can follow this protocol to readily obtain multiple GFP reporter iPSC lines that are useful for in vivo and in vitro imaging and cell isolation.

Una visualizzazione del protocollo è fornita in figura 2, con i periodi durante i quali iPSCs sono coltivate in mezzo diverso evidenziata da una verde per E8 o blu per NutriStem. E &#39;importante per trasfettare solo iPSCs di alta qualità; esaminare piatti della cultura in tutta la manutenzione ordinaria e verificare che le culture IPSC contengono colonie principalmente distinti recanti una morfologia ciottoli-simile (Figura 3A); cellule differenziate non dovrebbe occupare più del 1...

Watch this Scientific Journal Video about Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor at JoVE.com

Transfection, Selection, and Colony-picking of Human Induced Pluripotent Stem Cells TALEN-targeted with a GFP Gene into the AAVS1 Safe Harbor

TALEN-mediated gene editing at the safe harbor AAVS1 locus enables high-efficiency transgene addition in human iPSCs. This protocol describes the procedures for preparing iPSCs for TALEN and donor vector delivery, transfecting iPSCs, and selecting and isolating iPSC clones to achieve targeted integration of a GFP gene to generate reporter lines.

Transfection, selezione, e Colonia-picking di cellule staminali umane pluripotenti indotte TALEN-mirati con Gene GFP nel AAVS1 Safe Harbor

Targeted transgene addition can provide persistent gene expression while circumventing the gene silencing and insertional mutagenesis caused by viral ...

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Three-dimensional Quantification of Dendritic Spines from Pyramidal Neurons Derived from Human Induced Pluripotent Stem Cells

Quantificazione tridimensionale di spine dendritiche da piramidale neuroni derivate da cellule staminali umane pluripotenti indotte

Dendritic spines are small protrusions that correspond to the post-synaptic compartments of excitatory synapses in the central nervous system. They are ...

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Protocol for the Differentiation of Human Induced Pluripotent Stem Cells into Mixed Cultures of Neurons and Glia for Neurotoxicity Testing

Protocollo per la differenziazione delle cellule staminali pluripotenti indotte dall&#39;uomo in culture miste di neuroni e glia per la sperimentazione della neurotossicità

Human pluripotent stem cells can differentiate into various cell types that can be applied to human-based in vitro toxicity assays. One major advantage is ...

Differentiating Chondrocytes from Peripheral Blood-derived Human Induced Pluripotent Stem Cells

Differenziare i condrociti da cellule staminali pluripotenti indotte dall&#39;uomo derivato dal sangue

In this study, we used peripheral blood cells (PBCs) as seed cells to produce chondrocytes via induced pluripotent stem cells (iPSCs) in an ...

Using Human Induced Pluripotent Stem Cell-derived Hepatocyte-like Cells for Drug Discovery

Utilizzando umani indotti dell'epatocita-come di derivati da cellule staminali pluripotenti per scoperta della droga

The ability to differentiate human induced pluripotent stem cells (iPSCs) into hepatocyte-like cells (HLCs) provides new opportunities to study inborn ...

A Familial Hypercholesterolemia Human Liver Chimeric Mouse Model Using Induced Pluripotent Stem Cell-derived Hepatocytes

Un modello di Mouse chimerici del fegato umano di ipercolesterolemia familiare facendo uso degli epatociti derivati da cellule staminali pluripotenti indotte

Familial hypercholesterolemia (FH) is mostly caused by low-density lipoprotein receptor (LDLR) mutations and results in an increased risk of early-onset ...

RNA-based Reprogramming of Human Primary Fibroblasts into Induced Pluripotent Stem Cells

RNA-ha basato riprogrammazione dei fibroblasti primari umani in cellule staminali pluripotenti indotte

Induced pluripotent stem cells (iPSCs) have proven to be a valuable tool to study human development and disease. Further advancing iPSCs as a regenerative ...

Efficient Differentiation of Human Pluripotent Stem Cells into Liver Cells

Differenziazione efficiente delle cellule staminali pluripotenti umane nelle cellule del fegato

The liver detoxifies harmful substances, secretes vital proteins, and executes key metabolic activities, thus sustaining life. Consequently, liver ...

Generation of 3D Skin Organoid from Cord Blood-derived Induced Pluripotent Stem Cells

Generazione di pelle 3D Organoid dal cordone emoderivati indotta da cellule staminali pluripotenti indotte

The skin is the body&#8217;s largest organ and has many functions. The skin acts as a physical barrier and protector of the body and regulates bodily ...