Derivation of a diploid human embryonic stem cell line from a mononuclear zygote
BACKGROUND: IVF occasionally produces aneuploid zygotes with one or three pronuclei (PN). Routinely, these zygotes are discarded. The aim of this work was to establish human embryonic stem cell (hESC) lines from blastocysts resulting from abnormal fertilization. METHODS: Abnormally fertilized zygote...
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| Veröffentlicht in: | Human reproduction (Oxford) 2004-03, Vol.19 (3), p.670-675 |
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| creator | Suss‐Toby, Edith Gerecht‐Nir, S. Amit, M. Manor, D. Itskovitz‐Eldor, J. |
| description | BACKGROUND: IVF occasionally produces aneuploid zygotes with one or three pronuclei (PN). Routinely, these zygotes are discarded. The aim of this work was to establish human embryonic stem cell (hESC) lines from blastocysts resulting from abnormal fertilization. METHODS: Abnormally fertilized zygotes were cultured to the blastocyst stage and, following zona pellucida digestion, zona‐free blastocysts were placed on a mouse feeder layer. Culture of hESCs was carried out as described earlier. RESULTS: Six out of the nine developing blastocysts attached to the feeder layer. One hESC line, originating from a mononuclear zygote following ICSI, was successfully derived. This line displayed typical phenotype and embryonic surface markers, and exhibited the potential to develop into all three embryonic germ layers both in vitro (by embryoid body formation) and in vivo (teratoma generation). Genetic examination revealed normal diploid karyotype and heterozygotic appearance for metachromatic leukodystrophy (MLD). CONCLUSION: This method, which requires neither immuno nor mechanical removal of the trophectoderm, may facilitate the derivation of hESC lines in general, and those from abnormal embryos in particular. Furthermore, it is shown that aneuploid zygotes can be used as a source for normal hESC derivation and hold the potential to generate aneuploid hESC lines for research purposes. |
| doi_str_mv | 10.1093/humrep/deh135 |
| format | Article |
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Routinely, these zygotes are discarded. The aim of this work was to establish human embryonic stem cell (hESC) lines from blastocysts resulting from abnormal fertilization. METHODS: Abnormally fertilized zygotes were cultured to the blastocyst stage and, following zona pellucida digestion, zona‐free blastocysts were placed on a mouse feeder layer. Culture of hESCs was carried out as described earlier. RESULTS: Six out of the nine developing blastocysts attached to the feeder layer. One hESC line, originating from a mononuclear zygote following ICSI, was successfully derived. This line displayed typical phenotype and embryonic surface markers, and exhibited the potential to develop into all three embryonic germ layers both in vitro (by embryoid body formation) and in vivo (teratoma generation). Genetic examination revealed normal diploid karyotype and heterozygotic appearance for metachromatic leukodystrophy (MLD). CONCLUSION: This method, which requires neither immuno nor mechanical removal of the trophectoderm, may facilitate the derivation of hESC lines in general, and those from abnormal embryos in particular. Furthermore, it is shown that aneuploid zygotes can be used as a source for normal hESC derivation and hold the potential to generate aneuploid hESC lines for research purposes.</description><identifier>ISSN: 0268-1161</identifier><identifier>ISSN: 1460-2350</identifier><identifier>EISSN: 1460-2350</identifier><identifier>DOI: 10.1093/humrep/deh135</identifier><identifier>PMID: 14998969</identifier><identifier>CODEN: HUREEE</identifier><language>eng</language><publisher>Oxford: Oxford University Press</publisher><subject>aneuploid zygote/derivation/differentiation/embryonic stem cells ; Animals ; Biological and medical sciences ; Cell Line ; Cell Nucleus - ultrastructure ; Cells, Cultured ; Diploidy ; Embryology: invertebrates and vertebrates. Teratology ; Fertilization in Vitro ; Fundamental and applied biological sciences. Psychology ; Heterozygote ; Hindlimb ; Humans ; In Vitro Techniques ; Karyotyping ; Leukodystrophy, Metachromatic - genetics ; Mice ; Muscle Neoplasms - etiology ; Muscle Neoplasms - pathology ; Stem Cell Transplantation ; Stem Cells - cytology ; Stem Cells - physiology ; Teratoma - etiology ; Teratoma - pathology ; Transplantation, Heterologous ; Zygote - cytology ; Zygote - ultrastructure</subject><ispartof>Human reproduction (Oxford), 2004-03, Vol.19 (3), p.670-675</ispartof><rights>European Society of Human Reproduction and Embryology 2004</rights><rights>2004 INIST-CNRS</rights><rights>Copyright Oxford University Press(England) Mar 2004</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c522t-7cb4ce4578c3f4341903716562b97a34161eab3ffde6e0783840dbca5ec507493</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,1584,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15663658$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14998969$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Suss‐Toby, Edith</creatorcontrib><creatorcontrib>Gerecht‐Nir, S.</creatorcontrib><creatorcontrib>Amit, M.</creatorcontrib><creatorcontrib>Manor, D.</creatorcontrib><creatorcontrib>Itskovitz‐Eldor, J.</creatorcontrib><title>Derivation of a diploid human embryonic stem cell line from a mononuclear zygote</title><title>Human reproduction (Oxford)</title><addtitle>Hum. Reprod</addtitle><addtitle>Hum. Reprod</addtitle><description>BACKGROUND: IVF occasionally produces aneuploid zygotes with one or three pronuclei (PN). Routinely, these zygotes are discarded. The aim of this work was to establish human embryonic stem cell (hESC) lines from blastocysts resulting from abnormal fertilization. METHODS: Abnormally fertilized zygotes were cultured to the blastocyst stage and, following zona pellucida digestion, zona‐free blastocysts were placed on a mouse feeder layer. Culture of hESCs was carried out as described earlier. RESULTS: Six out of the nine developing blastocysts attached to the feeder layer. One hESC line, originating from a mononuclear zygote following ICSI, was successfully derived. This line displayed typical phenotype and embryonic surface markers, and exhibited the potential to develop into all three embryonic germ layers both in vitro (by embryoid body formation) and in vivo (teratoma generation). Genetic examination revealed normal diploid karyotype and heterozygotic appearance for metachromatic leukodystrophy (MLD). CONCLUSION: This method, which requires neither immuno nor mechanical removal of the trophectoderm, may facilitate the derivation of hESC lines in general, and those from abnormal embryos in particular. Furthermore, it is shown that aneuploid zygotes can be used as a source for normal hESC derivation and hold the potential to generate aneuploid hESC lines for research purposes.</description><subject>aneuploid zygote/derivation/differentiation/embryonic stem cells</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Cell Line</subject><subject>Cell Nucleus - ultrastructure</subject><subject>Cells, Cultured</subject><subject>Diploidy</subject><subject>Embryology: invertebrates and vertebrates. Teratology</subject><subject>Fertilization in Vitro</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Heterozygote</subject><subject>Hindlimb</subject><subject>Humans</subject><subject>In Vitro Techniques</subject><subject>Karyotyping</subject><subject>Leukodystrophy, Metachromatic - genetics</subject><subject>Mice</subject><subject>Muscle Neoplasms - etiology</subject><subject>Muscle Neoplasms - pathology</subject><subject>Stem Cell Transplantation</subject><subject>Stem Cells - cytology</subject><subject>Stem Cells - physiology</subject><subject>Teratoma - etiology</subject><subject>Teratoma - pathology</subject><subject>Transplantation, Heterologous</subject><subject>Zygote - cytology</subject><subject>Zygote - ultrastructure</subject><issn>0268-1161</issn><issn>1460-2350</issn><issn>1460-2350</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkE1r3DAURUVpaSbTLrstohDIxolkWbK8LJOvwkAGpiEhGyHLz41S23IkO2Ty66vBJll29fTg6N7HQegbJSeUFOz0YWw99KcVPFDGP6AFzQRJUsbJR7QgqZAJpYIeoMMQHgmJTyk-owOaFYUsRLFAmzPw9lkP1nXY1VjjyvaNsxWOubrD0JZ-5zprcBigxQaaBje2A1x710a6dZ3rRtOA9vh198cN8AV9qnUT4Os8l-jm4vz36ipZX1_-Wv1cJ4an6ZDkpswMZDyXhtUZy2hBWE4FF2lZ5DrugoIuWV1XIIDkksmMVKXRHAwneVawJfox5fbePY0QBvXoRt_FSpVSKqUU8dMSJRNkvAvBQ616b1vtd4oStdenJn1q0hf573PoWLZQvdOzrwgczYAORje1152x4Z3jQjDB98XHE-fG_r-d8402Kn55g7X_q0TOcq6u7u7VdrNdp2e3VG3YP56Sluk</recordid><startdate>20040301</startdate><enddate>20040301</enddate><creator>Suss‐Toby, Edith</creator><creator>Gerecht‐Nir, S.</creator><creator>Amit, M.</creator><creator>Manor, D.</creator><creator>Itskovitz‐Eldor, J.</creator><general>Oxford University Press</general><general>Oxford Publishing Limited (England)</general><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>K9.</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>20040301</creationdate><title>Derivation of a diploid human embryonic stem cell line from a mononuclear zygote</title><author>Suss‐Toby, Edith ; Gerecht‐Nir, S. ; Amit, M. ; Manor, D. ; Itskovitz‐Eldor, J.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c522t-7cb4ce4578c3f4341903716562b97a34161eab3ffde6e0783840dbca5ec507493</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>aneuploid zygote/derivation/differentiation/embryonic stem cells</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Cell Line</topic><topic>Cell Nucleus - ultrastructure</topic><topic>Cells, Cultured</topic><topic>Diploidy</topic><topic>Embryology: invertebrates and vertebrates. Teratology</topic><topic>Fertilization in Vitro</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Heterozygote</topic><topic>Hindlimb</topic><topic>Humans</topic><topic>In Vitro Techniques</topic><topic>Karyotyping</topic><topic>Leukodystrophy, Metachromatic - genetics</topic><topic>Mice</topic><topic>Muscle Neoplasms - etiology</topic><topic>Muscle Neoplasms - pathology</topic><topic>Stem Cell Transplantation</topic><topic>Stem Cells - cytology</topic><topic>Stem Cells - physiology</topic><topic>Teratoma - etiology</topic><topic>Teratoma - pathology</topic><topic>Transplantation, Heterologous</topic><topic>Zygote - cytology</topic><topic>Zygote - ultrastructure</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Suss‐Toby, Edith</creatorcontrib><creatorcontrib>Gerecht‐Nir, S.</creatorcontrib><creatorcontrib>Amit, M.</creatorcontrib><creatorcontrib>Manor, D.</creatorcontrib><creatorcontrib>Itskovitz‐Eldor, J.</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Human reproduction (Oxford)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Suss‐Toby, Edith</au><au>Gerecht‐Nir, S.</au><au>Amit, M.</au><au>Manor, D.</au><au>Itskovitz‐Eldor, J.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Derivation of a diploid human embryonic stem cell line from a mononuclear zygote</atitle><jtitle>Human reproduction (Oxford)</jtitle><stitle>Hum. Reprod</stitle><addtitle>Hum. Reprod</addtitle><date>2004-03-01</date><risdate>2004</risdate><volume>19</volume><issue>3</issue><spage>670</spage><epage>675</epage><pages>670-675</pages><issn>0268-1161</issn><issn>1460-2350</issn><eissn>1460-2350</eissn><coden>HUREEE</coden><abstract>BACKGROUND: IVF occasionally produces aneuploid zygotes with one or three pronuclei (PN). Routinely, these zygotes are discarded. The aim of this work was to establish human embryonic stem cell (hESC) lines from blastocysts resulting from abnormal fertilization. METHODS: Abnormally fertilized zygotes were cultured to the blastocyst stage and, following zona pellucida digestion, zona‐free blastocysts were placed on a mouse feeder layer. Culture of hESCs was carried out as described earlier. RESULTS: Six out of the nine developing blastocysts attached to the feeder layer. One hESC line, originating from a mononuclear zygote following ICSI, was successfully derived. This line displayed typical phenotype and embryonic surface markers, and exhibited the potential to develop into all three embryonic germ layers both in vitro (by embryoid body formation) and in vivo (teratoma generation). Genetic examination revealed normal diploid karyotype and heterozygotic appearance for metachromatic leukodystrophy (MLD). CONCLUSION: This method, which requires neither immuno nor mechanical removal of the trophectoderm, may facilitate the derivation of hESC lines in general, and those from abnormal embryos in particular. Furthermore, it is shown that aneuploid zygotes can be used as a source for normal hESC derivation and hold the potential to generate aneuploid hESC lines for research purposes.</abstract><cop>Oxford</cop><pub>Oxford University Press</pub><pmid>14998969</pmid><doi>10.1093/humrep/deh135</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Oxford University Press Journals All Titles (1996-Current) |
| subjects | aneuploid zygote/derivation/differentiation/embryonic stem cells Animals Biological and medical sciences Cell Line Cell Nucleus - ultrastructure Cells, Cultured Diploidy Embryology: invertebrates and vertebrates. Teratology Fertilization in Vitro Fundamental and applied biological sciences. Psychology Heterozygote Hindlimb Humans In Vitro Techniques Karyotyping Leukodystrophy, Metachromatic - genetics Mice Muscle Neoplasms - etiology Muscle Neoplasms - pathology Stem Cell Transplantation Stem Cells - cytology Stem Cells - physiology Teratoma - etiology Teratoma - pathology Transplantation, Heterologous Zygote - cytology Zygote - ultrastructure |
| title | Derivation of a diploid human embryonic stem cell line from a mononuclear zygote |
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