Name: direct induction of hemogenic endothelium and blood by overexpression of transcription factors in human pluripotent stem cells
Uploaded: 2015-12-03T10:30:00
Description: Watch this Scientific Journal Video about Direct Induction of Hemogenic Endothelium and Blood by Overexpression of Transcription Factors in Human Pluripotent Stem Cells at JoVE.com

We thank Matt Raymond for editorial assistance. This work was supported by funds from the National Institute of Health (U01HL099773, R01HL116221, and P51 RR000167) and The Charlotte Geyer Foundation.

1. Virus Forberedelser og transkripsjonsfaktor Kombinasjoner <ol><li> Forbered løsninger med pSIN-EF1α lentiviral uttrykk plasmid som inneholder protein-kodende DNA for ETV2, GATA1, GATA2, TAL1 og LMO2 (tabell 1). </li><li> Måle konsentrasjonen og renheten av plasmid preparater som brukes for lentiviral produksjon ved å registrere UV-absorpsjon med et spektrofotometer ved 230 nm, 260 nm og 280 nm. Merk: DNA-preparater som viser A260 / 280 og A260 / 230-verdier større enn 1,8 er generelt an...

<ol>
	<li>Zape, J. P., Zovein, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemogenic+endothelium:+origins,+regulation,+and+implications+for+vascular+biology.">Hemogenic endothelium: origins, regulation, and implications for vascular biology.</a> Semin Cell Dev Biol. 22, 1036-1047 (2011).</li><li>Swiers, G., Rode, C., Azzoni, E., de Bruijn, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+short+history+of+hemogenic+endothelium.">A short history of hemogenic endothelium.</a> Blood Cells, Mol &amp; Dis. 51, 206-212 (2013).</li><li>Kennedy, M., 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=T+lymphocyte+potential+marks+the+emergence+of+definitive+hematopoietic+progenitors+in+human+pluripotent+stem+cell+differentiation+cultures.">T lymphocyte potential marks the emergence of definitive hematopoietic progenitors in human pluripotent stem cell differentiation cultures.</a> Cell Rep. 2, 1722-1735 (2012).</li><li>Wang, L., 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=Endothelial+and+hematopoietic+cell+fate+of+human+embryonic+stem+cells+originates+from+primitive+endothelium+with+hemangioblastic+properties.">Endothelial and hematopoietic cell fate of human embryonic stem cells originates from primitive endothelium with hemangioblastic properties.</a> Immunity. 21, 31-41 (2004).</li><li>Rafii, S., 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=Human+ESC-derived+hemogenic+endothelial+cells+undergo+distinct+waves+of+endothelial+to+hematopoietic+transition.">Human ESC-derived hemogenic endothelial cells undergo distinct waves of endothelial to hematopoietic transition.</a> Blood. 121 (5), 770-780 (2012).</li><li>Choi, K. 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=Identification+of+the+hemogenic+endothelial+progenitor+and+its+direct+precursor+in+human+pluripotent+stem+cell+differentiation+cultures.">Identification of the hemogenic endothelial progenitor and its direct precursor in human pluripotent stem cell differentiation cultures.</a> Cell Rep. 2, 553-567 (2012).</li><li>Vodyanik, M. A., Thomson, J. A., Slukvin, I. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Leukosialin+(CD43)+defines+hematopoietic+progenitors+in+human+embryonic+stem+cell+differentiation+cultures.">Leukosialin (CD43) defines hematopoietic progenitors in human embryonic stem cell differentiation cultures.</a> Blood. 108, 2095-2105 (2006).</li><li>Wang, C., 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=TGFbeta+inhibition+enhances+the+generation+of+hematopoietic+progenitors+from+human+ES+cell-derived+hemogenic+endothelial+cells+using+a+stepwise+strategy.">TGFbeta inhibition enhances the generation of hematopoietic progenitors from human ES cell-derived hemogenic endothelial cells using a stepwise strategy.</a> Cell Res. 22, 194-207 (2012).</li><li>Uenishi, G., 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=Tenascin+C+promotes+hematoendothelial+development+and+T+lymphoid+commitment+from+human+pluripotent+stem+cells+in+chemically+defined+conditions.">Tenascin C promotes hematoendothelial development and T lymphoid commitment from human pluripotent stem cells in chemically defined conditions.</a> Stem Cell Rep. 3, 1073-1084 (2014).</li><li>Elcheva, I., 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=Direct+induction+of+haematoendothelial+programs+in+human+pluripotent+stem+cells+by+transcriptional+regulators.">Direct induction of haematoendothelial programs in human pluripotent stem cells by transcriptional regulators.</a> Nat Commun. 5, 4372 (2014).</li><li>Tiscornia, G., Singer, O., Verma, I. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+and+purification+of+lentiviral+vectors.">Production and purification of lentiviral vectors.</a> Nat Protoc. 1, 241-245 (2006).</li><li>Vodyanik, M. A., Slukvin, I. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hematoendothelial+differentiation+of+human+embryonic+stem+cells.">Hematoendothelial differentiation of human embryonic stem cells.</a> Curr Protoc Cell Biol. Chapter 23, Unit 23.6 (2007).</li><li>Cao, F., 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=Comparison+of+gene-transfer+efficiency+in+human+embryonic+stem+cells.">Comparison of gene-transfer efficiency in human embryonic stem cells.</a> Mol Imgn and Biol. 12, 15-24 (2010).</li></ol>

Den ovenfor beskrevne metode for hematopoietisk differensiering av hPSCs ved overekspresjon av TFS, representerer en hurtig og effektiv metode for generering av HE og myeloide og erytho-magakaryocytic progenitorceller fra hESCs og iPSCs, for derved å tillate produksjon av opp til 30 millioner blodceller fra én million pluripotente stamceller 10. Denne metoden viste konsekvent differensiering i flere hESC og iPSCs linjer 10. Under differensiering av ETV2 og GATA2, GATA1 faktorer samt GATA2 og TAL1...

Den unike evnen av menneskelige pluripotente stamceller (hPSCs) å fornye seg selv og til å differensiere i celler av de tre bakterie lag, inkludert blod, gjøre dem et verdifullt verktøy for mekanistiske studier av blodkreft utvikling, modellering av blodsykdommer, narkotika screening, toksisitetsstudier, samt utvikling av mobilnettet terapier. Fordi bloddannelse i embryo inntektene fra hemogenic endotel (HE) gjennom en endothelial hematopoietisk overgang 1,2, vil genereringen av HE i kulturer være avgjørende for å studere de molekylære mekanismene som regulerer endotelial til hematopoetiske overgang og hematopoetisk spesifikasjon. Aktuelle fremgangsmåter for studier av HE er basert på induksjon av differensiering i hematoendothelial aggregater (EBS) med tillegg av hematopoetiske cytokiner 3-5, og coculture av hPSCs med hematopoiesis støttende stromaceller 6,7 eller i to-dimensjonale kulturer med ekstracellulære matriser og cytokines 8,9. Disse klassiske differensierings metoder er basert på innføring av eksterne signaler som virker på celleoverflaten og initiere kaskader av molekylære reaksjonsveier som til slutt fører til aktivering av transkripsjonen program førings hematoendothelial utvikling. Således effektiviteten av hPSCs differensiering i disse systemene er avhengig av en effektiv induksjon av disse signalene, signaltransduksjon til kjernen, og den resulterende aktivering av spesifikke transkripsjons regulatorer. I tillegg har studier av HE i konvensjonelle differensierings kulturer krever ytterligere trinn for isolering HE celler ved hjelp av cellesortering. Her beskriver vi en enkel protokoll for direkte induksjon av HE og blod ved overekspresjon av blodkreft transkripsjonsfaktorer. Denne metoden gjør det mulig for effektiv induksjon av HE i en skål og direkte observasjon av endotelial til hematopoetiske overgang uten behov for isolering av HE ved hjelp av en cellesortering tungvint prosedyre. Dannelse av HE og blod fra menneskelige pluripotente stamceller kan effektivt indusert av overekspresjon bare noen transkripsjonsfaktorer (TFS). Den optimale kombinasjonen av TFS som kan indusere robust pan-myeloid hematopoiesen fra hPSCs inkluderer ETV2 og GATA1 eller GATA2. I kontrast, kombinasjon av GATA2 og TAL1 induserer erythromegakaryocytopoiesis 10. Programmering hPSCs gjennom overekspresjon av disse faktorene skiller hPSCs direkte til VE-cad + CD43 - CD73 - han celler som gradvis erverve blodkreft fenotype definert av uttrykk for tidlig blodkreft markør CD43 7. Dette lentiviral basert metode for direkte programmering av human flerpotent stamceller Metoden er anvendelig for generering av HE og blodceller for mekanistiske undersøkelser, studier av endotelial til hematopoetiske overgang, og transkripsjonsregulering av hematopoetiske utvikling og spesifikasjon. Selv om cjeldende protokollen beskriver blod produksjon ved hjelp konstituerende uttrykk for transgener, lignende resultater kan oppnås ved hjelp av endret mRNA 10.

Table 1. Induction of hPSCs differentiation with trancription factors and analysis of hemogenic endothelium and blood cells.
<table>
	<tbody>
		<tr>
			<td>pSIN4-EF1a-ETV2-IRES-Puro&#160;</td>
			<td>Addgene</td>
			<td>Plasmid #61061</td>
			<td>Lentiviral Vector</td>
		</tr>
		<tr>
			<td>pSIN4-EF1a-GATA2-IRES-Puro&#160;</td>
			<td>Addgene</td>
			<td>Plasmid #61063</td>
			<td>Lentiviral Vector</td>
		</tr>
		<tr>
			<td>pSIN4-EF1a-GATA1-IRES-Puro&#160;</td>
			<td>Addgene</td>
			<td>Plasmid #61062</td>
			<td>Lentiviral Vector</td>
		</tr>
		<tr>
			<td>pSIN4-EF1a-TAL1-IRES-Puro&#160;</td>
			<td>Addgene</td>
			<td>Plasmid #61062</td>
			<td>Lentiviral Vector</td>
		</tr>
		<tr>
			<td>pSIN4-EF1a-LMO2-IRES-Puro&#160;</td>
			<td>Addgene</td>
			<td>Plasmid #61064</td>
			<td>Lentiviral Vector</td>
		</tr>
		<tr>
			<td>Hexadimethrine bromide (Polybrene)&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>107689-10G</td>
			<td>Cationic polymer used to increase the efficiency of infection</td>
		</tr>
		<tr>
			<td>Y-27632 (Dihydrochloride) ROCK inhibitor</td>
			<td>STEMCELL Technologies</td>
			<td>72302</td>
			<td>RHO/ROCK pathway inhibitor Inhibits ROCK</td>
		</tr>
		<tr>
			<td rowspan="2">StemPro Accutase Cell Dissociation Reagent</td>
			<td rowspan="2"><a href="http://www.lifetechnologies.com/us/en/home/life-science/stem-cell-research/stem-cell-culture/stem-cell-research-misc/stempro-accutase.html">Life Technologies</a></td>
			<td rowspan="2">A11105-01</td>
			<td rowspan="2">Cell Dissociation Reagent</td>
		</tr>
		<tr>
		</tr>
		<tr>
			<td>Incomplete (growth factor- free) culture medium&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; mTeSR1 Custom formulation&#160;</td>
			<td>WiCell Research Institute (Madison, WI)</td>
			<td>MCF</td>
			<td><a name="RANGE!D12">Serum-free mTeSR1 medium without bFGF and TGFb&#160;</a></td>
		</tr>
		<tr>
			<td>human SCF</td>
			<td>Peprotech</td>
			<td>300-07</td>
			<td>Premium grade</td>
		</tr>
		<tr>
			<td>human TPO</td>
			<td>Peprotech</td>
			<td>300-18</td>
			<td>Research grade</td>
		</tr>
		<tr>
			<td>human FGF-basic</td>
			<td>Peprotech</td>
			<td>100-18B</td>
			<td>Premium grade</td>
		</tr>
		<tr>
			<td>CD144 (VE-cad) FITC</td>
			<td>BD Biosciences</td>
			<td>560411</td>
			<td>Endothelial marker (FACS)</td>
		</tr>
		<tr>
			<td>CD226 PE</td>
			<td>BD Biosciences</td>
			<td>338305</td>
			<td>Hematopoietic (FACS)</td>
		</tr>
		<tr>
			<td>CD43 PE</td>
			<td>BD Biosciences</td>
			<td>560199</td>
			<td>Hematopoietic (FACS)</td>
		</tr>
		<tr>
			<td>CD73 APC</td>
			<td>R&#38;D Systems</td>
			<td>FAB5795A</td>
			<td>Endothelial marker (FACS)</td>
		</tr>
		<tr>
			<td>CD45 APC</td>
			<td>BD Biosciences</td>
			<td>555485</td>
			<td>Hematopoietic (FACS)</td>
		</tr>
		<tr>
			<td>7AAD</td>
			<td>Life Technologies</td>
			<td>A1310</td>
			<td>Live/Dead assay (FACS)</td>
		</tr>
		<tr>
			<td>Paraformaldehyde&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>P6148-500G</td>
			<td>Cell fixation&#160;</td>
		</tr>
		<tr>
			<td>Triton X-100&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>T9284-500ML&#160;</td>
			<td>Permeabilization&#160;</td>
		</tr>
		<tr>
			<td>FBS</td>
			<td>Fisher Scientific</td>
			<td>SH3007003</td>
			<td>Fetal bovine serum</td>
		</tr>
		<tr>
			<td>Mouse anti-human CD43</td>
			<td>BD Biosciences</td>
			<td>551457</td>
			<td>Pure, primary antibody for Immunofluorescence (IF) staining</td>
		</tr>
		<tr>
			<td>Rabbit anti-human VE-cadherin</td>
			<td>BenderMedSystem</td>
			<td>BMS158</td>
			<td>Primary (IF)</td>
		</tr>
		<tr>
			<td>Anti-rabbit Alexa Fluor 488-conjugated</td>
			<td>JacksonResearch</td>
			<td>715-486-152</td>
			<td>Secondary (IF)</td>
		</tr>
		<tr>
			<td>Anti-mouse Alexa Fluor 594-conjugated</td>
			<td>JacksonResearch</td>
			<td>715-516-150&#160;</td>
			<td>Secondary (IF)</td>
		</tr>
		<tr>
			<td>DAPI nucleic acid stain</td>
			<td><a href="http://www.lifetechnologies.com/us/en/home/life-science/stem-cell-research/stem-cell-culture/stem-cell-research-misc/stempro-accutase.html">Life Technologies</a></td>
			<td>D1306&#160;</td>
			<td>Live/Dead assay (IF)</td>
		</tr>
		<tr>
			<td>Clonogenic medium MethoCult H4435 Enriched</td>
			<td>STEMCELL Technologies</td>
			<td>4435</td>
			<td>CFC-assay</td>
		</tr>
		<tr>
			<td>Wright Stain solution</td>
			<td>Sigma -Aldrich</td>
			<td>32857</td>
			<td>Staining cytospins</td>
		</tr>
	</tbody>
</table>

Table 2. hPSCs culture.
<table>
	<tbody>
		<tr>
			<td>Human Pluripotent Stem Cells (hPSCs)&#160;</td>
			<td>WiCell Research Institute (Madison, WI)</td>
			<td>hESCs (WA01, WA09) human Embryonic Stem cells; iPSCs (DF-19-9-7T, DF-4-3-7T) transgene-free induced Pluripotent Stem Cells</td>
			<td>hPSCs are able to self-renew and to differentiate into cells of three germ layers</td>
		</tr>
		<tr>
			<td>Complete serum-free medium for culture of hPSCs&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; mTeSR1 media&#160;&#160;</td>
			<td>WiCell Research Institute (Madison, WI)</td>
			<td>M500&#160;</td>
			<td>Serum-free&#160; medium with growth factors for feeder free culture of ESC/iPSCs</td>
		</tr>
		<tr>
			<td>Matrigel</td>
			<td>BD Biosciences/ Corning&#160;</td>
			<td>356234</td>
			<td>Matrix for maintenance of human ESC/iPSCs</td>
		</tr>
		<tr>
			<td>DMEM/F-12, powder&#160;</td>
			<td><a href="http://www.lifetechnologies.com/us/en/home/life-science/stem-cell-research/stem-cell-culture/stem-cell-research-misc/stempro-accutase.html">Life Technologies</a></td>
			<td>12500-062</td>
			<td>Basal Medium&#160;</td>
		</tr>
		<tr>
			<td>HyClone Dulbecco&#39;s PBS powder</td>
			<td>Fisher Scientific</td>
			<td>dSH30013.04</td>
			<td>PBS</td>
		</tr>
		<tr>
			<td rowspan="2">Dispase II, powder</td>
			<td rowspan="2"><a href="http://www.lifetechnologies.com/us/en/home/life-science/stem-cell-research/stem-cell-culture/stem-cell-research-misc/stempro-accutase.html">Life Technologies</a></td>
			<td rowspan="2">17105-041</td>
			<td rowspan="2">Neutral protease, Cell dissociation</td>
		</tr>
		<tr>
		</tr>
	</tbody>
</table>
Table 3. Primers for detection of virus genomic integration.
<table>
	<tbody>
		<tr>
			<td>Primer&#39;s Name</td>
			<td>Forward (Fwd)&#160; 5&#8217; --&#62;3&#8217;</td>
			<td>Reverse (Rev) 5&#8217;--&#62;3&#8217;</td>
			<td>Discription&#160;</td>
		</tr>
		<tr>
			<td>pSIN EF1a Fwd</td>
			<td>TTC CAT TTC AGG TGT CGT GA</td>
			<td>--</td>
			<td>EF1a promoter sequence</td>
		</tr>
		<tr>
			<td>GATA1 Rev</td>
			<td>--</td>
			<td>TCC CTG TAG TAG GCC AGT GC</td>
			<td>&#160;Coding Region&#160;</td>
		</tr>
		<tr>
			<td>GATA2 Rev</td>
			<td>--</td>
			<td>GGT TGG CAT AGT AGG GGT TG</td>
			<td>&#160;Coding Region&#160;</td>
		</tr>
		<tr>
			<td>TAL1 Rev</td>
			<td>--</td>
			<td>AGG CGG AGG ATC TCA TTC TT</td>
			<td>&#160;Coding Region&#160;</td>
		</tr>
		<tr>
			<td>LMO2 Rev</td>
			<td>--</td>
			<td>GGC CCA GTT TGT AGT AGA GGC</td>
			<td>&#160;Coding Region&#160;</td>
		</tr>
		<tr>
			<td>ETV2 Rev</td>
			<td>--</td>
			<td>GAA CTT CTG GGT GCA GTA AC&#160;</td>
			<td>&#160;Coding Region&#160;</td>
		</tr>
	</tbody>
</table>

direct induction of hemogenic endothelium and blood by overexpression of transcription factors in human pluripotent stem cells

Under utviklingen hematopoetiske celler oppstå fra en spesiell undergruppe av endotelceller, hemogenic endotel (HE). Modellering HE utvikling in vitro er vesentlig for mekanistiske undersøkelser av endotelial-hematopoetiske overgang og hematopoetisk spesifikasjon. Her beskriver vi en fremgangsmåte for effektiv induksjon av HE fra humane pluripotente stamceller (hPSCs) ved hjelp av overekspresjon av forskjellige sett av transkripsjonsfaktorer. Kombinasjonen av ETV2 og GATA1 eller GATA2 TFS blir brukt til å indusere HE med pan-myeloid potensiale, mens en kombinasjon av GATA2 og TAL1 transkripsjonsfaktorer gjør det mulig for produksjon av HE med erytroid og megakaryocytic potensial. Tilsetting av LMO2 til GATA2 og TAL1 kombinasjon vesentlig akselererer differensiering og øker erytroid og megakaryocytic celler produksjon. Denne metoden gir en effektiv og rask måte å HE induksjon fra hPSCs og tillater observasjon av endotelial-hematopoietic overgang i en kultur tallerken. Protokollen inkluderer hPSCs transduksjon prosedyrer og post-transduksjon analyse av HE og blodstamfedre.

Skjemaet på HE og blod induksjon fra hPSCs av overekspresjon av transkripsjonsfaktorer er vist i figur 1. ETV2 med GATA1 eller GATA2 kombinasjon induserer pan-myeloid hematopoiesen, mens en GATA2, TAL1 +/- LMO2 kombinasjon induserer hovedsakelig erythro-megakaryocytic hematopoiesen. Begge TF kombinasjoner direkte indusert HE celler som deretter forvandlet til blodstamfedre med en tydelig spekter av blodkreft differensiering. Differensiering av hPSCs fra pluripotent tilstand til blodproduserende HE tar ...

Watch this Scientific Journal Video about Direct Induction of Hemogenic Endothelium and Blood by Overexpression of Transcription Factors in Human Pluripotent Stem Cells at JoVE.com

Direct Induction of Hemogenic Endothelium and Blood by Overexpression of Transcription Factors in Human Pluripotent Stem Cells

This protocol describes the efficient induction of hemogenic endothelium and multipotential hematopoietic progenitors from human pluripotent stem cells via the forced expression of transcription factors.

Direkte Induksjon av Hemogenic endotelet og Blood av Overekspresjon av transkripsjonsfaktorer i Human Pluripotent stamceller

During development, hematopoietic cells arise from a specialized subset of endothelial cells, hemogenic endothelium (HE). Modeling HE development in vitro ...

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The specification of hemogenic endothelial cells from embryonic vascular endothelium occurs during brief developmental periods within distinct tissues, ...

Reprogramming Mouse Embryonic Fibroblasts with Transcription Factors to Induce a Hemogenic Program

Omprogrammering mus embryonale Fibroblaster med transkripsjonsfaktorer for å indusere en Hemogenic Program

This protocol details the induction of a hemogenic program in mouse embryonic fibroblasts (MEFs) via overexpression of transcription factors (TFs). We ...

Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors

Generasjon Integrerings--frie Utførte Pluripotent stamceller fra menneskelige perifere mononukleære blodceller Bruke episomale vektorer

Induced Pluripotent Stem Cells (iPSCs) hold great promise for disease modeling and regenerative therapies. We previously reported the use of Episomal ...

Directed Differentiation of Primitive and Definitive Hematopoietic Progenitors from Human Pluripotent Stem Cells

Rettet differensiering av Primitive og Definitive blodkreft Progenitors fra menneskelige Pluripotent stamceller

One of the major goals for regenerative medicine is the generation and maintenance of hematopoietic stem cells (HSCs) derived from human pluripotent stem ...

In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells

In Vitro Differentiation of Human Pluripotent Stem Cells into Trophoblastic Cells

In Vitro Differensiering av menneskelige pluripotent stamceller til trophoblastic Cells

The placenta is the first organ to develop during embryogenesis and is required for the survival of the developing embryo. The placenta is comprised of ...

Defined and Scalable Generation of Hepatocyte-like Cells from Human Pluripotent Stem Cells

Definert og Scalable Generation of hepatocytter lignende celler fra menneskelige Pluripotent stamceller

Human pluripotent stem cells (hPSCs) possess great value for biomedical research. hPSCs can be scaled and differentiated to all cell types found in the ...