Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos
Identifieur interne : 000B11 ( Istex/Corpus ); précédent : 000B10; suivant : 000B12Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos
Auteurs : Mieke Geens ; Ileana Mateizel ; Karen Sermon ; Martine De Rycke ; Claudia Spits ; Greet Cauffman ; Paul Devroey ; Herman Tournaye ; Inge Liebaers ; Hilde Van De VeldeSource :
- Human Reproduction [ 0268-1161 ] ; 2009-11.
Abstract
BACKGROUND Recently, we demonstrated that single blastomeres of a 4-cell stage human embryo are able to develop into blastocysts with inner cell mass and trophectoderm. To further investigate potency at the 4-cell stage, we aimed to derive pluripotent human embryonic stem cells (hESC) from single blastomeres. METHODS Four 4-cell stage embryos were split on Day 2 of preimplantation development and the 16 blastomeres were individually cultured in sequential medium. On Day 3 or 4, the blastomere-derived embryos were plated on inactivated mouse embryonic fibroblasts (MEFs). RESULTS Ten out of sixteen blastomere-derived morulae attached to the MEFs, and two produced an outgrowth. They were mechanically passaged onto fresh MEFs as described for blastocyst ICM-derived hESC, and shown to express the typical stemness markers by immunocytochemistry and/or RTPCR. In vivo pluripotency was confirmed by the presence of all three germ layers in the teratoma obtained after injection in immunodeficient mice. The first hESC line displays a mosaic normal/abnormal 46, XX, dup(7)(q33qter), del(18)(q23qter) karyotype. The second hESC line displays a normal 46, XY karyotype. CONCLUSION We report the successful derivation and characterization of two hESC lines from single blastomeres of four split 4-cell stage human embryos. These two hESC lines were derived from distinct embryos, proving that at least one of the 4-cell stage blastomeres is pluripotent.
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DOI: 10.1093/humrep/dep262
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<record><TEI wicri:istexFullTextTei="biblStruct"><teiHeader><fileDesc><titleStmt><title>Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos</title><author><name sortKey="Geens, Mieke" sort="Geens, Mieke" uniqKey="Geens M" first="Mieke" last="Geens">Mieke Geens</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Mateizel, Ileana" sort="Mateizel, Ileana" uniqKey="Mateizel I" first="Ileana" last="Mateizel">Ileana Mateizel</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Sermon, Karen" sort="Sermon, Karen" uniqKey="Sermon K" first="Karen" last="Sermon">Karen Sermon</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="De Rycke, Martine" sort="De Rycke, Martine" uniqKey="De Rycke M" first="Martine" last="De Rycke">Martine De Rycke</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Spits, Claudia" sort="Spits, Claudia" uniqKey="Spits C" first="Claudia" last="Spits">Claudia 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Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Tournaye, Herman" sort="Tournaye, Herman" uniqKey="Tournaye H" first="Herman" last="Tournaye">Herman Tournaye</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Liebaers, Inge" sort="Liebaers, Inge" uniqKey="Liebaers I" first="Inge" last="Liebaers">Inge Liebaers</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Van De Velde, Hilde" sort="Van De Velde, Hilde" uniqKey="Van De Velde H" first="Hilde" last="Van De Velde">Hilde Van De Velde</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: hilde.vandevelde@uzbrussel.be</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:7D197296D4CC1B1CA2DBDC4A6CEEC63C3FC1FB1C</idno><date when="2009" year="2009">2009</date><idno type="doi">10.1093/humrep/dep262</idno><idno type="url">https://api.istex.fr/document/7D197296D4CC1B1CA2DBDC4A6CEEC63C3FC1FB1C/fulltext/pdf</idno><idno type="wicri:Area/Istex/Corpus">000B11</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a">Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos</title><author><name sortKey="Geens, Mieke" sort="Geens, Mieke" uniqKey="Geens M" first="Mieke" last="Geens">Mieke Geens</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Mateizel, Ileana" sort="Mateizel, Ileana" uniqKey="Mateizel I" first="Ileana" last="Mateizel">Ileana Mateizel</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Sermon, Karen" sort="Sermon, Karen" uniqKey="Sermon K" first="Karen" last="Sermon">Karen Sermon</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="De Rycke, Martine" sort="De Rycke, Martine" uniqKey="De Rycke M" first="Martine" last="De Rycke">Martine De Rycke</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Spits, Claudia" sort="Spits, Claudia" uniqKey="Spits C" first="Claudia" last="Spits">Claudia Spits</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Cauffman, Greet" sort="Cauffman, Greet" uniqKey="Cauffman G" first="Greet" last="Cauffman">Greet Cauffman</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Devroey, Paul" sort="Devroey, Paul" uniqKey="Devroey P" first="Paul" last="Devroey">Paul Devroey</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Tournaye, Herman" sort="Tournaye, Herman" uniqKey="Tournaye H" first="Herman" last="Tournaye">Herman Tournaye</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Liebaers, Inge" sort="Liebaers, Inge" uniqKey="Liebaers I" first="Inge" last="Liebaers">Inge Liebaers</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation></author><author><name sortKey="Van De Velde, Hilde" sort="Van De Velde, Hilde" uniqKey="Van De Velde H" first="Hilde" last="Van De Velde">Hilde Van De Velde</name><affiliation><mods:affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: hilde.vandevelde@uzbrussel.be</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Human Reproduction</title><idno type="ISSN">0268-1161</idno><idno type="eISSN">1460-2350</idno><imprint><publisher>Oxford University Press</publisher><date type="published" when="2009-11">2009-11</date><biblScope unit="volume">24</biblScope><biblScope unit="issue">11</biblScope><biblScope unit="page" from="2709">2709</biblScope><biblScope unit="page" to="2717">2717</biblScope></imprint><idno type="ISSN">0268-1161</idno></series><idno type="istex">7D197296D4CC1B1CA2DBDC4A6CEEC63C3FC1FB1C</idno><idno type="DOI">10.1093/humrep/dep262</idno><idno type="ArticleID">dep262</idno></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0268-1161</idno></seriesStmt></fileDesc><profileDesc><textClass></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract">BACKGROUND Recently, we demonstrated that single blastomeres of a 4-cell stage human embryo are able to develop into blastocysts with inner cell mass and trophectoderm. To further investigate potency at the 4-cell stage, we aimed to derive pluripotent human embryonic stem cells (hESC) from single blastomeres. METHODS Four 4-cell stage embryos were split on Day 2 of preimplantation development and the 16 blastomeres were individually cultured in sequential medium. On Day 3 or 4, the blastomere-derived embryos were plated on inactivated mouse embryonic fibroblasts (MEFs). RESULTS Ten out of sixteen blastomere-derived morulae attached to the MEFs, and two produced an outgrowth. They were mechanically passaged onto fresh MEFs as described for blastocyst ICM-derived hESC, and shown to express the typical stemness markers by immunocytochemistry and/or RTPCR. In vivo pluripotency was confirmed by the presence of all three germ layers in the teratoma obtained after injection in immunodeficient mice. The first hESC line displays a mosaic normal/abnormal 46, XX, dup(7)(q33qter), del(18)(q23qter) karyotype. The second hESC line displays a normal 46, XY karyotype. CONCLUSION We report the successful derivation and characterization of two hESC lines from single blastomeres of four split 4-cell stage human embryos. These two hESC lines were derived from distinct embryos, proving that at least one of the 4-cell stage blastomeres is pluripotent.</div></front></TEI><istex><corpusName>oup</corpusName><author><json:item><name>Mieke Geens</name><affiliations><json:string>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string></affiliations></json:item><json:item><name>Ileana Mateizel</name><affiliations><json:string>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string></affiliations></json:item><json:item><name>Karen Sermon</name><affiliations><json:string>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string><json:string>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string></affiliations></json:item><json:item><name>Martine De Rycke</name><affiliations><json:string>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string><json:string>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string></affiliations></json:item><json:item><name>Claudia Spits</name><affiliations><json:string>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string></affiliations></json:item><json:item><name>Greet Cauffman</name><affiliations><json:string>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string><json:string>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string></affiliations></json:item><json:item><name>Paul Devroey</name><affiliations><json:string>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string><json:string>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string></affiliations></json:item><json:item><name>Herman Tournaye</name><affiliations><json:string>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string><json:string>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string></affiliations></json:item><json:item><name>Inge Liebaers</name><affiliations><json:string>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string><json:string>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string></affiliations></json:item><json:item><name>Hilde Van de Velde</name><affiliations><json:string>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string><json:string>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</json:string><json:string>E-mail: hilde.vandevelde@uzbrussel.be</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>human embryonic stem cell line</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>single blastomere</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>morula</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>4-cell stage human embryo</value></json:item></subject><articleId><json:string>dep262</json:string></articleId><language><json:string>eng</json:string></language><originalGenre><json:string>research-article</json:string></originalGenre><abstract>BACKGROUND Recently, we demonstrated that single blastomeres of a 4-cell stage human embryo are able to develop into blastocysts with inner cell mass and trophectoderm. To further investigate potency at the 4-cell stage, we aimed to derive pluripotent human embryonic stem cells (hESC) from single blastomeres. METHODS Four 4-cell stage embryos were split on Day 2 of preimplantation development and the 16 blastomeres were individually cultured in sequential medium. On Day 3 or 4, the blastomere-derived embryos were plated on inactivated mouse embryonic fibroblasts (MEFs). RESULTS Ten out of sixteen blastomere-derived morulae attached to the MEFs, and two produced an outgrowth. They were mechanically passaged onto fresh MEFs as described for blastocyst ICM-derived hESC, and shown to express the typical stemness markers by immunocytochemistry and/or RTPCR. In vivo pluripotency was confirmed by the presence of all three germ layers in the teratoma obtained after injection in immunodeficient mice. The first hESC line displays a mosaic normal/abnormal 46, XX, dup(7)(q33qter), del(18)(q23qter) karyotype. The second hESC line displays a normal 46, XY karyotype. CONCLUSION We report the successful derivation and characterization of two hESC lines from single blastomeres of four split 4-cell stage human embryos. These two hESC lines were derived from distinct embryos, proving that at least one of the 4-cell stage blastomeres is pluripotent.</abstract><qualityIndicators><score>8.372</score><pdfVersion>1.5</pdfVersion><pdfPageSize>612.283 x 790.866 pts</pdfPageSize><refBibsNative>true</refBibsNative><keywordCount>4</keywordCount><abstractCharCount>1453</abstractCharCount><pdfWordCount>4804</pdfWordCount><pdfCharCount>30546</pdfCharCount><pdfPageCount>9</pdfPageCount><abstractWordCount>214</abstractWordCount></qualityIndicators><title>Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos</title><genre><json:string>research-article</json:string></genre><host><volume>24</volume><publisherId><json:string>humrep</json:string></publisherId><pages><last>2717</last><first>2709</first></pages><issn><json:string>0268-1161</json:string></issn><issue>11</issue><genre><json:string>journal</json:string></genre><language><json:string>unknown</json:string></language><eissn><json:string>1460-2350</json:string></eissn><title>Human Reproduction</title></host><categories><wos><json:string>OBSTETRICS & GYNECOLOGY</json:string><json:string>REPRODUCTIVE BIOLOGY</json:string></wos></categories><publicationDate>2009</publicationDate><copyrightDate>2009</copyrightDate><doi><json:string>10.1093/humrep/dep262</json:string></doi><id>7D197296D4CC1B1CA2DBDC4A6CEEC63C3FC1FB1C</id><score>0.26487154</score><fulltext><json:item><original>true</original><mimetype>application/pdf</mimetype><extension>pdf</extension><uri>https://api.istex.fr/document/7D197296D4CC1B1CA2DBDC4A6CEEC63C3FC1FB1C/fulltext/pdf</uri></json:item><json:item><original>false</original><mimetype>application/zip</mimetype><extension>zip</extension><uri>https://api.istex.fr/document/7D197296D4CC1B1CA2DBDC4A6CEEC63C3FC1FB1C/fulltext/zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/document/7D197296D4CC1B1CA2DBDC4A6CEEC63C3FC1FB1C/fulltext/tei"><teiHeader><fileDesc><titleStmt><title level="a">Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>Oxford University Press</publisher><availability><p>The Author 2009. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissionsoxfordjournals.org</p></availability><date>2009-07-24</date></publicationStmt><sourceDesc><biblStruct type="inbook"><analytic><title level="a">Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos</title><author xml:id="author-1"><persName><forename type="first">Mieke</forename><surname>Geens</surname></persName><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation></author><author xml:id="author-2"><persName><forename type="first">Ileana</forename><surname>Mateizel</surname></persName><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation></author><author xml:id="author-3"><persName><forename type="first">Karen</forename><surname>Sermon</surname></persName><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation></author><author xml:id="author-4"><persName><forename type="first">Martine</forename><surname>De Rycke</surname></persName><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation></author><author xml:id="author-5"><persName><forename type="first">Claudia</forename><surname>Spits</surname></persName><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation></author><author xml:id="author-6"><persName><forename type="first">Greet</forename><surname>Cauffman</surname></persName><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation></author><author xml:id="author-7"><persName><forename type="first">Paul</forename><surname>Devroey</surname></persName><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation></author><author xml:id="author-8"><persName><forename type="first">Herman</forename><surname>Tournaye</surname></persName><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation></author><author xml:id="author-9"><persName><forename type="first">Inge</forename><surname>Liebaers</surname></persName><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation></author><author xml:id="author-10"><persName><forename type="first">Hilde</forename><surname>Van de Velde</surname></persName><email>hilde.vandevelde@uzbrussel.be</email><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation></author></analytic><monogr><title level="j">Human Reproduction</title><idno type="pISSN">0268-1161</idno><idno type="eISSN">1460-2350</idno><imprint><publisher>Oxford University Press</publisher><date type="published" when="2009-11"></date><biblScope unit="volume">24</biblScope><biblScope unit="issue">11</biblScope><biblScope unit="page" from="2709">2709</biblScope><biblScope unit="page" to="2717">2717</biblScope></imprint></monogr><idno type="istex">7D197296D4CC1B1CA2DBDC4A6CEEC63C3FC1FB1C</idno><idno type="DOI">10.1093/humrep/dep262</idno><idno type="ArticleID">dep262</idno></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>2009-07-24</date></creation><langUsage><language ident="en">en</language></langUsage><abstract><p>BACKGROUND Recently, we demonstrated that single blastomeres of a 4-cell stage human embryo are able to develop into blastocysts with inner cell mass and trophectoderm. To further investigate potency at the 4-cell stage, we aimed to derive pluripotent human embryonic stem cells (hESC) from single blastomeres. METHODS Four 4-cell stage embryos were split on Day 2 of preimplantation development and the 16 blastomeres were individually cultured in sequential medium. On Day 3 or 4, the blastomere-derived embryos were plated on inactivated mouse embryonic fibroblasts (MEFs). RESULTS Ten out of sixteen blastomere-derived morulae attached to the MEFs, and two produced an outgrowth. They were mechanically passaged onto fresh MEFs as described for blastocyst ICM-derived hESC, and shown to express the typical stemness markers by immunocytochemistry and/or RTPCR. In vivo pluripotency was confirmed by the presence of all three germ layers in the teratoma obtained after injection in immunodeficient mice. The first hESC line displays a mosaic normal/abnormal 46, XX, dup(7)(q33qter), del(18)(q23qter) karyotype. The second hESC line displays a normal 46, XY karyotype. CONCLUSION We report the successful derivation and characterization of two hESC lines from single blastomeres of four split 4-cell stage human embryos. These two hESC lines were derived from distinct embryos, proving that at least one of the 4-cell stage blastomeres is pluripotent.</p></abstract><textClass><keywords scheme="keyword"><list><head>keywords</head><item><term>human embryonic stem cell line</term></item><item><term>single blastomere</term></item><item><term>morula</term></item><item><term>4-cell stage human embryo</term></item></list></keywords></textClass></profileDesc><revisionDesc><change when="2009-07-24">Created</change><change when="2009-11">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><original>false</original><mimetype>text/plain</mimetype><extension>txt</extension><uri>https://api.istex.fr/document/7D197296D4CC1B1CA2DBDC4A6CEEC63C3FC1FB1C/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="corpus oup" wicri:toSee="no header"><istex:xmlDeclaration>version="1.0" encoding="utf-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//NLM//DTD Journal Publishing DTD v2.3 20070202//EN" URI="journalpublishing.dtd" name="istex:docType"></istex:docType><istex:document><article article-type="research-article"><front><journal-meta><journal-id journal-id-type="publisher-id">humrep</journal-id><journal-id journal-id-type="hwp">humrep</journal-id><journal-title>Human Reproduction</journal-title><issn pub-type="ppub">0268-1161</issn><issn pub-type="epub">1460-2350</issn><publisher><publisher-name>Oxford University Press</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="doi">10.1093/humrep/dep262</article-id><article-id pub-id-type="publisher-id">dep262</article-id><article-categories><subj-group subj-group-type="heading"><subject>ORIGINAL ARTICLES</subject><subj-group><subject>Embryology</subject></subj-group></subj-group></article-categories><title-group><article-title>Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Geens</surname><given-names>Mieke</given-names></name><xref ref-type="aff" rid="af1">1</xref></contrib><contrib contrib-type="author"><name><surname>Mateizel</surname><given-names>Ileana</given-names></name><xref ref-type="aff" rid="af1">1</xref></contrib><contrib contrib-type="author"><name><surname>Sermon</surname><given-names>Karen</given-names></name><xref ref-type="aff" rid="af1">1</xref><xref ref-type="aff" rid="af2">2</xref></contrib><contrib contrib-type="author"><name><surname>De Rycke</surname><given-names>Martine</given-names></name><xref ref-type="aff" rid="af1">1</xref><xref ref-type="aff" rid="af2">2</xref></contrib><contrib contrib-type="author"><name><surname>Spits</surname><given-names>Claudia</given-names></name><xref ref-type="aff" rid="af1">1</xref></contrib><contrib contrib-type="author"><name><surname>Cauffman</surname><given-names>Greet</given-names></name><xref ref-type="aff" rid="af1">1</xref><xref ref-type="aff" rid="af3">3</xref></contrib><contrib contrib-type="author"><name><surname>Devroey</surname><given-names>Paul</given-names></name><xref ref-type="aff" rid="af1">1</xref><xref ref-type="aff" rid="af3">3</xref></contrib><contrib contrib-type="author"><name><surname>Tournaye</surname><given-names>Herman</given-names></name><xref ref-type="aff" rid="af1">1</xref><xref ref-type="aff" rid="af3">3</xref></contrib><contrib contrib-type="author"><name><surname>Liebaers</surname><given-names>Inge</given-names></name><xref ref-type="aff" rid="af1">1</xref><xref ref-type="aff" rid="af2">2</xref></contrib><contrib contrib-type="author"><name><surname>Van de Velde</surname><given-names>Hilde</given-names></name><xref ref-type="aff" rid="af1">1</xref><xref ref-type="aff" rid="af3">3</xref><xref ref-type="corresp" rid="cor1">4</xref></contrib></contrib-group><aff id="af1"><label>1</label><addr-line>Department of Embryology and Genetics (EMGE)</addr-line>, <institution>Vrije Universiteit Brussel (VUB)</institution>, <addr-line>Laarbeeklaan 101, 1090 Brussels</addr-line>, <country>Belgium</country></aff><aff id="af2"><label>2</label><institution>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel)</institution>, <addr-line>Laarbeeklaan 101, 1090 Brussels</addr-line>, <country>Belgium</country></aff><aff id="af3"><label>3</label><institution>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel)</institution>, <addr-line>Laarbeeklaan 101, 1090 Brussels</addr-line>, <country>Belgium</country></aff><author-notes><corresp id="cor1"><label>4</label>Correspondence address. Tel: <phone>+32-24776690</phone>; <email>hilde.vandevelde@uzbrussel.be</email></corresp></author-notes><pub-date pub-type="ppub"><month>11</month><year>2009</year></pub-date><pub-date pub-type="epub"><day>24</day><month>7</month><year>2009</year></pub-date><volume>24</volume><issue>11</issue><fpage>2709</fpage><lpage>2717</lpage><history><date date-type="received"><day>9</day><month>7</month><year>2008</year></date><date date-type="rev-recd"><day>11</day><month>6</month><year>2009</year></date><date date-type="accepted"><day>18</day><month>6</month><year>2009</year></date></history><copyright-statement>© The Author 2009. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oxfordjournals.org</copyright-statement><copyright-year>2009</copyright-year><license license-type="creative-commons" xlink:href="http://creativecommons.org/licenses/by-nc/2.0/uk/"><p>The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed: the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given: if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative word this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org</p></license><abstract><sec><title>BACKGROUND</title><p>Recently, we demonstrated that single blastomeres of a 4-cell stage human embryo are able to develop into blastocysts with inner cell mass and trophectoderm. To further investigate potency at the 4-cell stage, we aimed to derive pluripotent human embryonic stem cells (hESC) from single blastomeres.</p></sec><sec><title>METHODS</title><p>Four 4-cell stage embryos were split on Day 2 of preimplantation development and the 16 blastomeres were individually cultured in sequential medium. On Day 3 or 4, the blastomere-derived embryos were plated on inactivated mouse embryonic fibroblasts (MEFs).</p></sec><sec><title>RESULTS</title><p>Ten out of sixteen blastomere-derived morulae attached to the MEFs, and two produced an outgrowth. They were mechanically passaged onto fresh MEFs as described for blastocyst ICM-derived hESC, and shown to express the typical stemness markers by immunocytochemistry and/or RT–PCR. I<italic>n vivo</italic> pluripotency was confirmed by the presence of all three germ layers in the teratoma obtained after injection in immunodeficient mice. The first hESC line displays a mosaic normal/abnormal 46, XX, dup(7)(q33qter), del(18)(q23qter) karyotype. The second hESC line displays a normal 46, XY karyotype.</p></sec><sec><title>CONCLUSION</title><p>We report the successful derivation and characterization of two hESC lines from single blastomeres of four split 4-cell stage human embryos. These two hESC lines were derived from distinct embryos, proving that at least one of the 4-cell stage blastomeres is pluripotent.</p></sec></abstract><kwd-group><kwd>human embryonic stem cell line</kwd><kwd>single blastomere</kwd><kwd>morula</kwd><kwd>4-cell stage human embryo</kwd></kwd-group></article-meta></front></article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo><title>Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos</title></titleInfo><titleInfo type="alternative" contentType="CDATA"><title>Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos</title></titleInfo><name type="personal"><namePart type="given">Mieke</namePart><namePart type="family">Geens</namePart><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Ileana</namePart><namePart type="family">Mateizel</namePart><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Karen</namePart><namePart type="family">Sermon</namePart><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Martine</namePart><namePart type="family">De Rycke</namePart><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Claudia</namePart><namePart type="family">Spits</namePart><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Greet</namePart><namePart type="family">Cauffman</namePart><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Paul</namePart><namePart type="family">Devroey</namePart><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Herman</namePart><namePart type="family">Tournaye</namePart><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Inge</namePart><namePart type="family">Liebaers</namePart><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Medical Genetics (CMG), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Hilde</namePart><namePart type="family">Van de Velde</namePart><affiliation>Department of Embryology and Genetics (EMGE), Vrije Universiteit Brussel (VUB), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>Centre for Reproductive Medicine (CRM), Universitair Ziekenhuis Brussel (UZ Brussel), Laarbeeklaan 101, 1090 Brussels, Belgium</affiliation><affiliation>E-mail: hilde.vandevelde@uzbrussel.be</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="research-article"></genre><originInfo><publisher>Oxford University Press</publisher><dateIssued encoding="w3cdtf">2009-11</dateIssued><dateCreated encoding="w3cdtf">2009-07-24</dateCreated><copyrightDate encoding="w3cdtf">2009</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><physicalDescription><internetMediaType>text/html</internetMediaType></physicalDescription><abstract>BACKGROUND Recently, we demonstrated that single blastomeres of a 4-cell stage human embryo are able to develop into blastocysts with inner cell mass and trophectoderm. To further investigate potency at the 4-cell stage, we aimed to derive pluripotent human embryonic stem cells (hESC) from single blastomeres. METHODS Four 4-cell stage embryos were split on Day 2 of preimplantation development and the 16 blastomeres were individually cultured in sequential medium. On Day 3 or 4, the blastomere-derived embryos were plated on inactivated mouse embryonic fibroblasts (MEFs). RESULTS Ten out of sixteen blastomere-derived morulae attached to the MEFs, and two produced an outgrowth. They were mechanically passaged onto fresh MEFs as described for blastocyst ICM-derived hESC, and shown to express the typical stemness markers by immunocytochemistry and/or RTPCR. In vivo pluripotency was confirmed by the presence of all three germ layers in the teratoma obtained after injection in immunodeficient mice. The first hESC line displays a mosaic normal/abnormal 46, XX, dup(7)(q33qter), del(18)(q23qter) karyotype. The second hESC line displays a normal 46, XY karyotype. CONCLUSION We report the successful derivation and characterization of two hESC lines from single blastomeres of four split 4-cell stage human embryos. These two hESC lines were derived from distinct embryos, proving that at least one of the 4-cell stage blastomeres is pluripotent.</abstract><subject><genre>keywords</genre><topic>human embryonic stem cell line</topic><topic>single blastomere</topic><topic>morula</topic><topic>4-cell stage human embryo</topic></subject><relatedItem type="host"><titleInfo><title>Human Reproduction</title></titleInfo><genre type="journal">journal</genre><identifier type="ISSN">0268-1161</identifier><identifier type="eISSN">1460-2350</identifier><identifier type="PublisherID">humrep</identifier><identifier type="PublisherID-hwp">humrep</identifier><part><date>2009</date><detail type="volume"><caption>vol.</caption><number>24</number></detail><detail type="issue"><caption>no.</caption><number>11</number></detail><extent unit="pages"><start>2709</start><end>2717</end></extent></part></relatedItem><identifier type="istex">7D197296D4CC1B1CA2DBDC4A6CEEC63C3FC1FB1C</identifier><identifier type="DOI">10.1093/humrep/dep262</identifier><identifier type="ArticleID">dep262</identifier><accessCondition type="use and reproduction" contentType="copyright">The Author 2009. 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|texte= Human embryonic stem cell lines derived from single blastomeres of two 4-cell stage embryos
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