Name: efficient ips cell generation from blood using episomes and hdac inhibitors
Uploaded: 2014-10-28T14:00:00
Description: Watch this Scientific Journal Video about Efficient iPS Cell Generation from Blood Using Episomes and HDAC Inhibitors at JoVE.com

Forfatterne ønsker at anerkende følgende tilskud fra NIH til at støtte denne forskning: K08DK082783 (AR), P30DK56465 (BTS), U01HL099993 (BTS), T32HL00715036 (SKS), og K12HL0806406 (SKS); og JP McCarthy Foundation (AR).

Skriftligt informeret samtykke, som er godkendt af de Institutional Review Boards af Fred Hutchinson Cancer Research Center og Children &quot;s Hospital i Philadelphia, blev opnået fra patienter før indsamlingen perifere blodprøver. Alle institutionelle retningslinier blev observeret. Alle dyreforsøg, herunder MEF generation og teratom dannelse blev godkendt af Institutional Animal Care og brug Udvalg. 1. Ficoll Adskillelse af PBMC&#39;er og Udvidelse af erytroblaster - Dag 0 <ol><li> ...

<ol>
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L., Warren, N., Sugden, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+replication+of+plasmids+derived+from+Epstein-Barr+virus+in+various+mammalian+cells.">Stable replication of plasmids derived from Epstein-Barr virus in various mammalian cells.</a> Nature. 313 (6005), 812-815 (1985).</li><li>Ware, C. B., 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=Histone+deacetylase+inhibition+elicits+an+evolutionarily+conserved+self-renewal+program+in+embryonic+stem+cells.">Histone deacetylase inhibition elicits an evolutionarily conserved self-renewal program in embryonic stem cells.</a> Cell Stem Cell. 4 (4), 359-369 (2009).</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=Butyrate+greatly+enhances+derivation+of+human+induced+pluripotent+stem+cells+by+promoting+epigenetic+remodeling+and+the+expression+of+pluripotency-associated+genes.">Butyrate greatly enhances derivation of human induced pluripotent stem cells by promoting epigenetic remodeling and the expression of pluripotency-associated genes.</a> Stem Cells. 28 (4), 713-720 (2010).</li><li>Azuara, V., 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=Chromatin+signatures+of+pluripotent+cell+lines.">Chromatin signatures of pluripotent cell lines.</a> Nature Cell Biology. 8 (5), 532-538 (2006).</li><li>Sommer, A. 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For vellykkede iPSC generation, når du bruger denne protokol, der er flere vigtige forbehold, der bør overvejes. Under erythroblast ekspansionsfasen, bør medierne ændre tidsplanen følges nøje, da afvigelser kan føre til ineffektiv stimulering af målet progenitorcellepopulation og en lavere effektivitet iPSC generation. Det er vigtigt at gøre ny ekspansion medium med frisk dexamethason med hver medier forandringer; og QBSF-60 basismedium og dexamethason bør beskyttes mod lys under opbevaring. Under nucleofectio...

iPSCs stammer fra somatiske væv via ektopisk ekspression af et minimalt sæt af pluripotens gener. Denne teknik blev oprindeligt påvist ved retroviral transduktion af humane fibroblaster med OCT4, SOX2, KLF4 og cMYC, som er stærkt udtrykt i den pluripotente tilstand 1. Disse forbigående udtrykt &quot;omprogrammering faktorer&quot; ændre target cellens epigenetisk landskab og genekspressionsprofil analog med humane embryonale stamceller 2. Når den er oprettet, kan iPSCs potentielt differentieres i nogen vævstyper til yderligere undersøgelse. Således, at de er lovende til anvendelse i regenerativ medicin, sygdomsmodeller og genterapianvendelser. Imidlertid forstyrre genomet med integrere vira har potentialet til at ændre endogene genekspression indflydelse cellulære fænotype, og i sidste ende forspænde videnskabelige resultater. Derudover kan tilfældige virale integrationer føre til skadelig cellulær effekter, herunder muligheden for malign transformation 3 eller re-ekspression af onkogene transgener 4. Fremtidige kliniske anvendelser vil kræve ikke-integrerende iPSC generation. Episomer, som er ekstra kromosomale cirkulære DNA-molekyler, tilbyder en strategi til at generere omkostningseffektive, integration-fri iPSCs 5. Kombinationen af episomale vektorer vist i tabel 1 udtrykke omprogrammering faktorer OCT4, SOX2, KLF4, MYCL og LIN28A. Den pCXLE_hOCT3 / 4-shp53-F plasmid indeholder også et p53 shRNA til midlertidig undertrykkelse af TP53 at forbedre cellulær omprogrammering 6. Den replikationsdeficient pCXWB-EBNA1 vektor fremmer opformering af omprogrammering faktorer og øget omprogrammering effektivitet ved at give en forbigående stigning i EBNA1 udtryk 7. Den pCXLE_EGFP plasmid kan tilsættes til nucleofection blandingen med henblik på determining transfektionseffektiviteten eller cellesortering applikationer. Med undtagelse af p CXWB-EBNA1 de episomale plasmider, der anvendes i denne protokol indeholder den Epstein-Barr virus af viral replikation og EBNA1-genet, som medierer replikation og deling af episom under deling af værtscellen 8. De episomer spontant tabt med successive iPSC ekspansion 7. Subkloning og karakterisering af iPSCs med episomalvektor tab, som kan udledes af tab af eGFP udtryk, kan føre til helt integration gratis iPSCs for fremtidige kliniske anvendelser. Iboende i processen iPSC generation er undertrykkelse af linjespecifikke gener og genaktivering af pluripotens-associerede gener. Regulering af genekspression forekommer på flere niveauer i kernen, herunder ændringer af DNA og chromatin at tillade transkriptionsfaktorer, regulatoriske DNA-elementer, og RNA-polymerase adgang til målrettetgener. Ombygning af epigenetiske landskab via globale kromatin modifikationer er en nøglekomponent til at re-ekspression af pluripotens genetiske program. En specifik chromatin modifikation, der er vigtig i regulering af genekspression er acetylering af histoner på bestemte lysinrester, som giver adgang til målgener gennem nedsat spænding af histon-DNA spole. HDAC-hæmmere er små molekyler, der har vist sig at forbedre iPSC omprogrammering og embryonale selvfornyelse 9,10, sandsynligvis på grund af støtte acetyleret tilstand 11. Den nedenfor beskrevne protokol, tilpasset fra en forudgående offentliggørelse hjælp integrere lentivira 12, giver en trin-for-trin metode til optimeret iPSC generation fra perifert blod ved hjælp episomer og HDAC-hæmmere. De HDAC inhibitorkoncentrationer anvendt her, er halvdelen af dem, der er beskrevet af Ware et al. 9 og har rutinemæssigt ført til en 2 gange stigning i fuldt omprogrammerede IPSC kolonier end standardepisomale omprogrammeringsbeslutninger protokoller uden HDAC-hæmmere. Dette niveau af omprogrammering er på niveau med den effektivitet, vi observerer med lentivirale metoder. Ved anvendelse af denne protokol, har vi effektivt genereret iPSC fra enkeltpersoner så gammel som 87 år gammel.

<table border="0" cellpadding="0" cellspacing="0" width="1100">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">Name of Reagent/ Equipment</td>
			<td style="width:151px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:503px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">Ficoll-Paque PLUS</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:119px;">45-001-750</td>
			<td>Store at 4&#176;C. Warm to room temperature before use.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">DPBS</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">14190-250</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">RPMI 1640</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">11875-093</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">fetal bovine serum (FBS)</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">10437028</td>
			<td>Store at -20&#176;C until needed. Thaw, aliquot, store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">dimethyl sulfoxide (DMSO)</td>
			<td style="width:151px;">Sigma-Aldrich</td>
			<td style="width:119px;">154938</td>
			<td>Store at room temperature.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">QBSF-60 serum-free medium</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:119px;">50-983-234</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">penicillin/streptomycin</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">15140122</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">Cell Line Nucleofector Kit V</td>
			<td style="width:151px;">Lonza</td>
			<td style="width:119px;">VCA-1003</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">2% gelatin solution</td>
			<td style="width:151px;">Sigma-Aldrich</td>
			<td style="width:119px;">G1393</td>
			<td>Store at 4&#176;C, liquify in 37&#176;C water bath before use.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">Hank&#39;s Balanced Saline Solution (HBSS)</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">14175-103</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:329px;">Dulbecco&#39;s Modified Eagle Medium (DMEM)</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">11965-092</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">Iscove&#39;s Modified Dulbecco&#39;s Medium (IMDM)</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">12440-061</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">L-glutamine, 200 mM</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">25030-081</td>
			<td>Aliquot, freeze at -20&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">non-essential amino acids (NEAA), 100X</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">11140-050</td>
			<td>Store at 4&#176;C, away from light.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">DMEM/Ham&#39;s F12, 1:1</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:119px;">SH30023.02</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">KnockOut Serum Replacement</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">10828-028</td>
			<td>Aliquot, freeze at -20&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">sodium pyruvate, 100 mM</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">11360-070</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">sodium bicarbonate, 7.5%</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">25080094</td>
			<td>Store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">basic fibroblast growth factor (bFGF)</td>
			<td style="width:151px;">Life Technologies</td>
			<td style="width:119px;">PHG0263</td>
			<td>Make 10 mg/mL stock in DPBS, aliquot, freeze at -20&#176;C. Once thawed, store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">sodium butyrate&#160;</td>
			<td style="width:151px;">Sigma-Aldrich</td>
			<td style="width:119px;">B5887-1G</td>
			<td>Make 2000X stock (400 mM) in DPBS, aliquot, freeze at -20&#176;C. Once thawed, store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">hES Cell Cloning &#38; Recovery Supplement</td>
			<td style="width:151px;">Stemgent</td>
			<td style="width:119px;">01-0014-500</td>
			<td>Store at -20&#176;C until needed. Once thawed, store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">ESC-qualified BD matrigel</td>
			<td style="width:151px;">BD Biosciences</td>
			<td style="width:119px;">35-4277</td>
			<td>Thaw overnight on ice at 4&#176;C, aliquot into pre-chilled tubes using pre-chilled pipette tips. Store at -20&#176;C until needed. Thaw at 4&#176;C, use immediately.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:329px;">StemSpan SFEM&#160;</td>
			<td style="width:151px;">STEMCELL Technologies</td>
			<td style="width:119px;">9650</td>
			<td>Aliquot, freeze at -20&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">ascorbic acid, powdered</td>
			<td style="width:151px;">Sigma-Aldrich</td>
			<td style="width:119px;">A4403-100MG</td>
			<td>Make 5 mg/mL stock in DPBS, sterile filter, store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">recombinant human stem cell factor (SCF)</td>
			<td style="width:151px;">R &#38; D Systems</td>
			<td style="width:119px;">255-SC-010</td>
			<td>Make 100 &#956;g/mL stock in SFEM, aliquot, freeze at -20&#176;C. Once thawed, store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">recombinant human interleukin 3 (IL-3)</td>
			<td style="width:151px;">R &#38; D Systems</td>
			<td style="width:119px;">203-IL-010</td>
			<td>Make 100 &#956;g/mL stock in SFEM, aliquot, freeze at -20&#176;C. Once thawed, store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">erythropoietin (EPO)</td>
			<td style="width:151px;">R &#38; D Systems</td>
			<td style="width:119px;">287-TC-500</td>
			<td>Make 1000 U/mL stock in SFEM, aliquot, freeze at -20&#176;C. Once thawed, store at 4&#176;C.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:329px;">recombinant human insulin-like growth factor 1 (IGF-1)</td>
			<td style="width:151px;">R &#38; D Systems</td>
			<td style="width:119px;">291-G1-200</td>
			<td>Make 100 &#956;g/mL stock in SFEM, aliquot, freeze at -20&#176;C. Once thawed, store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">&#946;-mercaptoethanol</td>
			<td style="width:151px;">Sigma-Aldrich</td>
			<td style="width:119px;">M7522</td>
			<td>Make 1000X stock (100 mM) in DPBS.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">dexamethasone</td>
			<td style="width:151px;">Sigma-Aldrich</td>
			<td style="width:119px;">D4902-25MG</td>
			<td>Make 50X stock (50 &#956;M) in DPBS, sterile filter, store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">SAHA (vorinostat)</td>
			<td style="width:151px;">Cayman Chemical</td>
			<td style="width:119px;">149647-78-9</td>
			<td>Make 2000X stock (400 mM) in DPBS, aliquot, freeze at -20&#176;C. Once thawed, store at 4&#176;C.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">12 well tissue culture plate</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:119px;">08-772-29</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">15 mL conical tube</td>
			<td style="width:151px;">Sarstedt</td>
			<td style="width:119px;">62553002</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">1.5 mL Eppendorf tube</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:119px;">05-408-129</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">6 well tissue culture plate</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:119px;">08-772-1B</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">35 mm tisue culture plates</td>
			<td style="width:151px;">BD Biosciences</td>
			<td style="width:119px;">353001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">10 mL disposable serological pipettes</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:119px;">13-675-20</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">5 mL disposable serological pipettes</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:119px;">13-675-22</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">2 mL disposable serological pipettes</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:119px;">13-675-17</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">20 &#956;L pipette tips, barrier tips</td>
			<td style="width:151px;">Genessee</td>
			<td style="width:119px;">24-404</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">glass Pasteur pipettes</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:119px;">13-678-20D</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">pipette aid</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:119px;">13-681-15</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">pCXLE-hOCT3/4-shp53-F</td>
			<td style="width:151px;">Addgene</td>
			<td style="width:119px;">27077</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">pCXLE-hSK</td>
			<td style="width:151px;">Addgene</td>
			<td style="width:119px;">27078</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">pCXLE-hUL</td>
			<td style="width:151px;">Addgene</td>
			<td style="width:119px;">27080</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">pCXLE-EGFP</td>
			<td style="width:151px;">Addgene</td>
			<td style="width:119px;">27082</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:329px;">pCXWB-EBNA1</td>
			<td style="width:151px;">Addgene</td>
			<td style="width:119px;">37624</td>
			<td></td>
		</tr>
	</tbody>
</table>

efficient ips cell generation from blood using episomes and hdac inhibitors

Dette håndskrift illustrerer en protokol for effektivt at skabe integration uden menneskeskabte pluripotente stamceller (iPSCs) fra perifert blod ved hjælp af episomale plasmider og histon deacetylase (HDAC) inhibitorer. Fordelene ved denne fremgangsmåde omfatter: (1) anvendelse af en minimal mængde af perifert blod som udgangsmateriale; (2) nonintegrating omprogrammering vektorer; (3) en omkostningseffektiv metode til at generere vektor gratis iPSCs; (4) en enkelt transfektion; og (5) anvendelse af små molekyler for at lette epigenetisk omprogrammering. Kort fortalt perifere mononukleære blodceller (PBMC&#39;er) isoleret fra rutinemæssige flebotomi prøver og derefter dyrket i definerede vækstfaktorer til opnåelse af en meget proliferativ erythrocyt progenitorcellepopulationen der er bemærkelsesværdigt modtagelig for omprogrammering. Nonintegrating, nontransmissible episomale plasmider, der udtrykker OCT4, SOX2, KLF4, MYCL, LIN28A og en p53 kort hårnål (sh) RNA er Introduced i de afledte erythroblaster via en enkelt nucleofection. Cotransfektion af et episom der udtrykker forstærket grønt fluorescerende protein (eGFP) giver mulighed for nem identifikation af transficerede celler. En separat replikationsdefekt plasmid, der udtrykker Epstein-Barr-nukleare antigen 1 (EBNA1) også sættes til reaktionsblandingen for forøget ekspression af episomale proteiner. Transficerede celler udplades derefter på et lag af bestrålet muse embryonale fibroblaster (iMEFs) for fortsat omprogrammering. Så snart iPSC-lignende kolonier vises omkring tolv dage efter nucleofection er HDAC-hæmmere til mediet for at lette epigenetisk remodeling. Vi har fundet, at inddragelsen af ​​HDAC-hæmmere rutinemæssigt øger generation af fuldt omprogrammerede IPSC kolonier med 2 gange. Når IPSC kolonier udviser typisk humane embryonale stamceller (hESC) morfologi, er de forsigtigt overført til individuelle iMEF-coatede vævskulturplader for fortsat vækst og ekspansion.

Tre dage efter nucleofection og før udpladning af celler på nucleofected iMEFs bør effektiviteten af vellykket nucleofection estimeres ved fluorescensmikroskopi for eGFP. Figur 1 viser en typisk nucleofection eksperiment med ca. 5-10% af den totale cellepopulation, der udtrykker eGFP. Omprogrammeres IPSC kolonier vil begynde at dukke cirka to uger efter nucleofection. Kolonierne er generelt cirkulær med veldefinerede grænser og kan identificeres ved karakteristiske embr...

Watch this Scientific Journal Video about Efficient iPS Cell Generation from Blood Using Episomes and HDAC Inhibitors at JoVE.com

Efficient iPS Cell Generation from Blood Using Episomes and HDAC Inhibitors

Her beskriver vi en protokol til generering af menneskeskabte pluripotente stamceller fra perifert blod ved hjælp af et episom baseret omprogrammering strategi og histondeacetylaseinhibitorer.

Effektiv iPS Cell Generation fra Blood Brug episomer og HDAC-hæmmere

This manuscript illustrates a protocol for efficiently creating integration-free human induced pluripotent stem cells (iPSCs) from peripheral blood using ...

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MicroRNA Expression Profiles of Human iPS Cells, Retinal Pigment Epithelium Derived From iPS, and Fetal Retinal Pigment Epithelium

MicroRNA Expression Profiler af menneskelige iPS celler, retinale pigmentepitel arvet fra iPS og Føtal retinale pigmentepitel

The objective of this report is to describe the protocols for comparing the microRNA (miRNA) profiles of human induced-pluripotent stem (iPS) cells, ...