Targeting a GFP reporter gene to the MIXL1 locus of human embryonic stem cells identifies human primitive streak–like cells and enables isolation of primitive hematopoietic precursors
Differentiating human embryonic stem cells (HESCs) represent an experimental platform for establishing the relationships between the earliest lineages that emerge during human development. Here we report the targeted insertion in HESCs of sequences encoding green fluorescent protein (GFP) into the l...
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| Veröffentlicht in: | Blood 2008-02, Vol.111 (4), p.1876-1884 |
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| container_issue | 4 |
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| container_title | Blood |
| container_volume | 111 |
| creator | Davis, Richard P. Ng, Elizabeth S. Costa, Magdaline Mossman, Anna K. Sourris, Koula Elefanty, Andrew G. Stanley, Edouard G. |
| description | Differentiating human embryonic stem cells (HESCs) represent an experimental platform for establishing the relationships between the earliest lineages that emerge during human development. Here we report the targeted insertion in HESCs of sequences encoding green fluorescent protein (GFP) into the locus of MIXL1, a gene transiently expressed in the primitive streak during embryogenesis.1,2 GFP fluorescence in MIXL1GFP/w HESCs differentiated in the presence of BMP4 reported the expression of MIXL1, permitting the identification of viable human primitive streak-like cells. The use of GFP as a reporter for MIXL1 combined with cell surface staining for platelet-derived growth factor receptor alpha (PDGFRα) enabled the isolation of a cell population that was highly enriched in primitive hematopoietic precursors, the earliest derivatives of the primitive streak. These experiments demonstrate the utility of MIXL1GFP/w HESCs for analyzing the previously inaccessible events surrounding the development of human primitive streak-like cells and their subsequent commitment to hematopoiesis. |
| doi_str_mv | 10.1182/blood-2007-06-093609 |
| format | Article |
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Here we report the targeted insertion in HESCs of sequences encoding green fluorescent protein (GFP) into the locus of MIXL1, a gene transiently expressed in the primitive streak during embryogenesis.1,2 GFP fluorescence in MIXL1GFP/w HESCs differentiated in the presence of BMP4 reported the expression of MIXL1, permitting the identification of viable human primitive streak-like cells. The use of GFP as a reporter for MIXL1 combined with cell surface staining for platelet-derived growth factor receptor alpha (PDGFRα) enabled the isolation of a cell population that was highly enriched in primitive hematopoietic precursors, the earliest derivatives of the primitive streak. These experiments demonstrate the utility of MIXL1GFP/w HESCs for analyzing the previously inaccessible events surrounding the development of human primitive streak-like cells and their subsequent commitment to hematopoiesis.</description><identifier>ISSN: 0006-4971</identifier><identifier>EISSN: 1528-0020</identifier><identifier>DOI: 10.1182/blood-2007-06-093609</identifier><identifier>PMID: 18032708</identifier><language>eng</language><publisher>Washington, DC: Elsevier Inc</publisher><subject>Animals ; Biological and medical sciences ; Embryonic Development ; Embryonic Stem Cells - cytology ; Embryonic Stem Cells - physiology ; Flow Cytometry ; Genes, Reporter ; Green Fluorescent Proteins - genetics ; Hematologic and hematopoietic diseases ; Hematopoietic Stem Cells - cytology ; Hematopoietic Stem Cells - physiology ; Homeodomain Proteins - genetics ; Humans ; Medical sciences ; Models, Animal</subject><ispartof>Blood, 2008-02, Vol.111 (4), p.1876-1884</ispartof><rights>2008 American Society of Hematology</rights><rights>2008 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c502t-75e05f7dc1548ec65bb474ff4b052465620f1b90cd43e5ea7e00797f0ac994f43</citedby><cites>FETCH-LOGICAL-c502t-75e05f7dc1548ec65bb474ff4b052465620f1b90cd43e5ea7e00797f0ac994f43</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=20121907$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18032708$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Davis, Richard P.</creatorcontrib><creatorcontrib>Ng, Elizabeth S.</creatorcontrib><creatorcontrib>Costa, Magdaline</creatorcontrib><creatorcontrib>Mossman, Anna K.</creatorcontrib><creatorcontrib>Sourris, Koula</creatorcontrib><creatorcontrib>Elefanty, Andrew G.</creatorcontrib><creatorcontrib>Stanley, Edouard G.</creatorcontrib><title>Targeting a GFP reporter gene to the MIXL1 locus of human embryonic stem cells identifies human primitive streak–like cells and enables isolation of primitive hematopoietic precursors</title><title>Blood</title><addtitle>Blood</addtitle><description>Differentiating human embryonic stem cells (HESCs) represent an experimental platform for establishing the relationships between the earliest lineages that emerge during human development. Here we report the targeted insertion in HESCs of sequences encoding green fluorescent protein (GFP) into the locus of MIXL1, a gene transiently expressed in the primitive streak during embryogenesis.1,2 GFP fluorescence in MIXL1GFP/w HESCs differentiated in the presence of BMP4 reported the expression of MIXL1, permitting the identification of viable human primitive streak-like cells. The use of GFP as a reporter for MIXL1 combined with cell surface staining for platelet-derived growth factor receptor alpha (PDGFRα) enabled the isolation of a cell population that was highly enriched in primitive hematopoietic precursors, the earliest derivatives of the primitive streak. These experiments demonstrate the utility of MIXL1GFP/w HESCs for analyzing the previously inaccessible events surrounding the development of human primitive streak-like cells and their subsequent commitment to hematopoiesis.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Embryonic Development</subject><subject>Embryonic Stem Cells - cytology</subject><subject>Embryonic Stem Cells - physiology</subject><subject>Flow Cytometry</subject><subject>Genes, Reporter</subject><subject>Green Fluorescent Proteins - genetics</subject><subject>Hematologic and hematopoietic diseases</subject><subject>Hematopoietic Stem Cells - cytology</subject><subject>Hematopoietic Stem Cells - physiology</subject><subject>Homeodomain Proteins - genetics</subject><subject>Humans</subject><subject>Medical sciences</subject><subject>Models, Animal</subject><issn>0006-4971</issn><issn>1528-0020</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc1u1DAUhS0EotPCGyDkDewC147zt0FCFS2VBsGiSOwsx7meMU3swXYqdcc78DR9HZ4Eh4nojpUl-zvX555DyAsGbxhr-dt-9H4oOEBTQF1AV9bQPSIbVvG2AODwmGwA8ovoGnZCTmP8DsBEyaun5IS1UPIG2g25v1Zhh8m6HVX08uILDXjwIWGgO3RIk6dpj_TT1bcto6PXc6Te0P08KUdx6sOdd1bTmHCiGscxUjugS9ZYjCt1CHayyd5ipgKqm98_f432BldcuYGiU_2YeRv9qJL1bvniQbbHSSV_8Da71Pke9RyiD_EZeWLUGPH5ep6Rrxcfrs8_FtvPl1fn77eFroCnoqkQKtMMmlWiRV1XfS8aYYzooeKirmoOhvUd6EGUWKFqMCfaNQaU7jphRHlGXh_nHoL_MWNMcrJxca8c-jnKBngrcpYZFEdQBx9jQCOXJVS4kwzkUpn8W5lcKpNQy2NlWfZynT_3Ew4PorWjDLxaARW1Gk1QTtv4j-PAOOugydy7I4c5jVuLQUZt0WkcbA4tycHb_zv5A4lgugk</recordid><startdate>20080215</startdate><enddate>20080215</enddate><creator>Davis, Richard P.</creator><creator>Ng, Elizabeth S.</creator><creator>Costa, Magdaline</creator><creator>Mossman, Anna K.</creator><creator>Sourris, Koula</creator><creator>Elefanty, Andrew G.</creator><creator>Stanley, Edouard G.</creator><general>Elsevier Inc</general><general>The Americain Society of Hematology</general><scope>6I.</scope><scope>AAFTH</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>7X8</scope></search><sort><creationdate>20080215</creationdate><title>Targeting a GFP reporter gene to the MIXL1 locus of human embryonic stem cells identifies human primitive streak–like cells and enables isolation of primitive hematopoietic precursors</title><author>Davis, Richard P. ; Ng, Elizabeth S. ; Costa, Magdaline ; Mossman, Anna K. ; Sourris, Koula ; Elefanty, Andrew G. ; Stanley, Edouard G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c502t-75e05f7dc1548ec65bb474ff4b052465620f1b90cd43e5ea7e00797f0ac994f43</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Embryonic Development</topic><topic>Embryonic Stem Cells - cytology</topic><topic>Embryonic Stem Cells - physiology</topic><topic>Flow Cytometry</topic><topic>Genes, Reporter</topic><topic>Green Fluorescent Proteins - genetics</topic><topic>Hematologic and hematopoietic diseases</topic><topic>Hematopoietic Stem Cells - cytology</topic><topic>Hematopoietic Stem Cells - physiology</topic><topic>Homeodomain Proteins - genetics</topic><topic>Humans</topic><topic>Medical sciences</topic><topic>Models, Animal</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Davis, Richard P.</creatorcontrib><creatorcontrib>Ng, Elizabeth S.</creatorcontrib><creatorcontrib>Costa, Magdaline</creatorcontrib><creatorcontrib>Mossman, Anna K.</creatorcontrib><creatorcontrib>Sourris, Koula</creatorcontrib><creatorcontrib>Elefanty, Andrew G.</creatorcontrib><creatorcontrib>Stanley, Edouard G.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</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>MEDLINE - Academic</collection><jtitle>Blood</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Davis, Richard P.</au><au>Ng, Elizabeth S.</au><au>Costa, Magdaline</au><au>Mossman, Anna K.</au><au>Sourris, Koula</au><au>Elefanty, Andrew G.</au><au>Stanley, Edouard G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Targeting a GFP reporter gene to the MIXL1 locus of human embryonic stem cells identifies human primitive streak–like cells and enables isolation of primitive hematopoietic precursors</atitle><jtitle>Blood</jtitle><addtitle>Blood</addtitle><date>2008-02-15</date><risdate>2008</risdate><volume>111</volume><issue>4</issue><spage>1876</spage><epage>1884</epage><pages>1876-1884</pages><issn>0006-4971</issn><eissn>1528-0020</eissn><abstract>Differentiating human embryonic stem cells (HESCs) represent an experimental platform for establishing the relationships between the earliest lineages that emerge during human development. Here we report the targeted insertion in HESCs of sequences encoding green fluorescent protein (GFP) into the locus of MIXL1, a gene transiently expressed in the primitive streak during embryogenesis.1,2 GFP fluorescence in MIXL1GFP/w HESCs differentiated in the presence of BMP4 reported the expression of MIXL1, permitting the identification of viable human primitive streak-like cells. The use of GFP as a reporter for MIXL1 combined with cell surface staining for platelet-derived growth factor receptor alpha (PDGFRα) enabled the isolation of a cell population that was highly enriched in primitive hematopoietic precursors, the earliest derivatives of the primitive streak. These experiments demonstrate the utility of MIXL1GFP/w HESCs for analyzing the previously inaccessible events surrounding the development of human primitive streak-like cells and their subsequent commitment to hematopoiesis.</abstract><cop>Washington, DC</cop><pub>Elsevier Inc</pub><pmid>18032708</pmid><doi>10.1182/blood-2007-06-093609</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| subjects | Animals Biological and medical sciences Embryonic Development Embryonic Stem Cells - cytology Embryonic Stem Cells - physiology Flow Cytometry Genes, Reporter Green Fluorescent Proteins - genetics Hematologic and hematopoietic diseases Hematopoietic Stem Cells - cytology Hematopoietic Stem Cells - physiology Homeodomain Proteins - genetics Humans Medical sciences Models, Animal |
| title | Targeting a GFP reporter gene to the MIXL1 locus of human embryonic stem cells identifies human primitive streak–like cells and enables isolation of primitive hematopoietic precursors |
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