Neural differentiation, selection and transcriptomic profiling of human neuromesodermal progenitor-like cells in vitro
Robust protocols for directed differentiation of human pluripotent cells are required to determine whether mechanisms operating in model organisms are relevant to our own development. Recent work in vertebrate embryos has identified neuromesodermal progenitors as a bipotent cell population that cont...
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Veröffentlicht in: | Development (Cambridge) 2018-07, Vol.145 (16) |
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creator | Verrier, Laure Davidson, Lindsay Gierliński, Marek Dady, Alwyn Storey, Kate G |
description | Robust protocols for directed differentiation of human pluripotent cells are required to determine whether mechanisms operating in model organisms are relevant to our own development. Recent work in vertebrate embryos has identified neuromesodermal progenitors as a bipotent cell population that contributes to paraxial mesoderm and spinal cord. However, precise protocols for
differentiation of human spinal cord progenitors are lacking. Informed by signalling in amniote embryos, we show here that transient dual-SMAD inhibition, together with retinoic acid (dSMADi-RA), provides rapid and reproducible induction of human spinal cord progenitors from neuromesodermal progenitor-like cells. Using CRISPR-Cas9 to engineer human embryonic stem cells with a GFP-reporter for neuromesodermal progenitor-associated gene
we facilitate selection of this cell population. RNA-sequencing was then used to identify human and conserved neuromesodermal progenitor transcriptional signatures, to validate this differentiation protocol and to reveal new pathways/processes in human neural differentiation. This optimised protocol, novel reporter line and transcriptomic data are useful resources with which to dissect molecular mechanisms regulating human spinal cord generation and allow the scaling-up of distinct cell populations for global analyses, including proteomic, biochemical and chromatin interrogation. |
doi_str_mv | 10.1242/dev.166215 |
format | Article |
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differentiation of human spinal cord progenitors are lacking. Informed by signalling in amniote embryos, we show here that transient dual-SMAD inhibition, together with retinoic acid (dSMADi-RA), provides rapid and reproducible induction of human spinal cord progenitors from neuromesodermal progenitor-like cells. Using CRISPR-Cas9 to engineer human embryonic stem cells with a GFP-reporter for neuromesodermal progenitor-associated gene
we facilitate selection of this cell population. RNA-sequencing was then used to identify human and conserved neuromesodermal progenitor transcriptional signatures, to validate this differentiation protocol and to reveal new pathways/processes in human neural differentiation. This optimised protocol, novel reporter line and transcriptomic data are useful resources with which to dissect molecular mechanisms regulating human spinal cord generation and allow the scaling-up of distinct cell populations for global analyses, including proteomic, biochemical and chromatin interrogation.</description><identifier>ISSN: 0950-1991</identifier><identifier>EISSN: 1477-9129</identifier><identifier>DOI: 10.1242/dev.166215</identifier><identifier>PMID: 29899136</identifier><language>eng</language><publisher>England: The Company of Biologists Ltd</publisher><subject>Animals ; Body Patterning - genetics ; Cell Differentiation - genetics ; Cell Lineage - genetics ; Cells, Cultured ; Embryo, Mammalian ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Human Development ; Humans ; Mesoderm - cytology ; Mesoderm - embryology ; Mesoderm - physiology ; Mice ; Neural Stem Cells - cytology ; Neural Stem Cells - physiology ; Neurogenesis - genetics ; Neurogenesis - physiology ; Spinal Cord - cytology ; Spinal Cord - physiology ; Stem Cells - cytology ; Stem Cells - physiology</subject><ispartof>Development (Cambridge), 2018-07, Vol.145 (16)</ispartof><rights>2018. Published by The Company of Biologists Ltd.</rights><rights>2018. Published by The Company of Biologists Ltd 2018</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><orcidid>0000-0003-3506-1287</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29899136$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Verrier, Laure</creatorcontrib><creatorcontrib>Davidson, Lindsay</creatorcontrib><creatorcontrib>Gierliński, Marek</creatorcontrib><creatorcontrib>Dady, Alwyn</creatorcontrib><creatorcontrib>Storey, Kate G</creatorcontrib><title>Neural differentiation, selection and transcriptomic profiling of human neuromesodermal progenitor-like cells in vitro</title><title>Development (Cambridge)</title><addtitle>Development</addtitle><description>Robust protocols for directed differentiation of human pluripotent cells are required to determine whether mechanisms operating in model organisms are relevant to our own development. Recent work in vertebrate embryos has identified neuromesodermal progenitors as a bipotent cell population that contributes to paraxial mesoderm and spinal cord. However, precise protocols for
differentiation of human spinal cord progenitors are lacking. Informed by signalling in amniote embryos, we show here that transient dual-SMAD inhibition, together with retinoic acid (dSMADi-RA), provides rapid and reproducible induction of human spinal cord progenitors from neuromesodermal progenitor-like cells. Using CRISPR-Cas9 to engineer human embryonic stem cells with a GFP-reporter for neuromesodermal progenitor-associated gene
we facilitate selection of this cell population. RNA-sequencing was then used to identify human and conserved neuromesodermal progenitor transcriptional signatures, to validate this differentiation protocol and to reveal new pathways/processes in human neural differentiation. This optimised protocol, novel reporter line and transcriptomic data are useful resources with which to dissect molecular mechanisms regulating human spinal cord generation and allow the scaling-up of distinct cell populations for global analyses, including proteomic, biochemical and chromatin interrogation.</description><subject>Animals</subject><subject>Body Patterning - genetics</subject><subject>Cell Differentiation - genetics</subject><subject>Cell Lineage - genetics</subject><subject>Cells, Cultured</subject><subject>Embryo, Mammalian</subject><subject>Gene Expression Profiling</subject><subject>Gene Expression Regulation, Developmental</subject><subject>Human Development</subject><subject>Humans</subject><subject>Mesoderm - cytology</subject><subject>Mesoderm - embryology</subject><subject>Mesoderm - physiology</subject><subject>Mice</subject><subject>Neural Stem Cells - cytology</subject><subject>Neural Stem Cells - physiology</subject><subject>Neurogenesis - genetics</subject><subject>Neurogenesis - physiology</subject><subject>Spinal Cord - cytology</subject><subject>Spinal Cord - physiology</subject><subject>Stem Cells - cytology</subject><subject>Stem Cells - physiology</subject><issn>0950-1991</issn><issn>1477-9129</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkE1PwzAMhiMEYmNw4QegHDlQSNIkbS5IaOJLmuAC5ypt3S3QJiVpK_HvycRAcLEt-30fy0bolJJLyji7qmG6pFIyKvbQnPIsSxRlah_NiRIkoUrRGToK4Y0QksosO0QzpvLYTeUcTU8wet3i2jQNeLCD0YNx9gIHaKHalljbGg9e21B50w-uMxXuvWtMa-wauwZvxk5bbCPHdRBcDb6LwChZgzWD80lr3gFX0LYBG4snM3h3jA4a3QY42eUFer27fVk-JKvn-8flzSrpmZRDIhqueKVZrfJKEF5y0WQaZElYXQHLtc6V4LnIGM-5UBUljNG65DHQTEdLukDX39x-LDuIJhsvaYvem077z8JpU_yfWLMp1m4qZPys4CwCzncA7z5GCEPRmbC9RVtwYygYEULSjPM8Ss_-7vpd8vPs9AtveYSO</recordid><startdate>20180712</startdate><enddate>20180712</enddate><creator>Verrier, Laure</creator><creator>Davidson, Lindsay</creator><creator>Gierliński, Marek</creator><creator>Dady, Alwyn</creator><creator>Storey, Kate G</creator><general>The Company of Biologists Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-3506-1287</orcidid></search><sort><creationdate>20180712</creationdate><title>Neural differentiation, selection and transcriptomic profiling of human neuromesodermal progenitor-like cells in vitro</title><author>Verrier, Laure ; Davidson, Lindsay ; Gierliński, Marek ; Dady, Alwyn ; Storey, Kate G</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p266t-5f494ca2d98c504b45f7ae6b02dce28aa8954857248459c10221db421d17a8c53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Animals</topic><topic>Body Patterning - genetics</topic><topic>Cell Differentiation - genetics</topic><topic>Cell Lineage - genetics</topic><topic>Cells, Cultured</topic><topic>Embryo, Mammalian</topic><topic>Gene Expression Profiling</topic><topic>Gene Expression Regulation, Developmental</topic><topic>Human Development</topic><topic>Humans</topic><topic>Mesoderm - cytology</topic><topic>Mesoderm - embryology</topic><topic>Mesoderm - physiology</topic><topic>Mice</topic><topic>Neural Stem Cells - cytology</topic><topic>Neural Stem Cells - physiology</topic><topic>Neurogenesis - genetics</topic><topic>Neurogenesis - physiology</topic><topic>Spinal Cord - cytology</topic><topic>Spinal Cord - physiology</topic><topic>Stem Cells - cytology</topic><topic>Stem Cells - physiology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Verrier, Laure</creatorcontrib><creatorcontrib>Davidson, Lindsay</creatorcontrib><creatorcontrib>Gierliński, Marek</creatorcontrib><creatorcontrib>Dady, Alwyn</creatorcontrib><creatorcontrib>Storey, Kate G</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Development (Cambridge)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Verrier, Laure</au><au>Davidson, Lindsay</au><au>Gierliński, Marek</au><au>Dady, Alwyn</au><au>Storey, Kate G</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Neural differentiation, selection and transcriptomic profiling of human neuromesodermal progenitor-like cells in vitro</atitle><jtitle>Development (Cambridge)</jtitle><addtitle>Development</addtitle><date>2018-07-12</date><risdate>2018</risdate><volume>145</volume><issue>16</issue><issn>0950-1991</issn><eissn>1477-9129</eissn><abstract>Robust protocols for directed differentiation of human pluripotent cells are required to determine whether mechanisms operating in model organisms are relevant to our own development. Recent work in vertebrate embryos has identified neuromesodermal progenitors as a bipotent cell population that contributes to paraxial mesoderm and spinal cord. However, precise protocols for
differentiation of human spinal cord progenitors are lacking. Informed by signalling in amniote embryos, we show here that transient dual-SMAD inhibition, together with retinoic acid (dSMADi-RA), provides rapid and reproducible induction of human spinal cord progenitors from neuromesodermal progenitor-like cells. Using CRISPR-Cas9 to engineer human embryonic stem cells with a GFP-reporter for neuromesodermal progenitor-associated gene
we facilitate selection of this cell population. RNA-sequencing was then used to identify human and conserved neuromesodermal progenitor transcriptional signatures, to validate this differentiation protocol and to reveal new pathways/processes in human neural differentiation. This optimised protocol, novel reporter line and transcriptomic data are useful resources with which to dissect molecular mechanisms regulating human spinal cord generation and allow the scaling-up of distinct cell populations for global analyses, including proteomic, biochemical and chromatin interrogation.</abstract><cop>England</cop><pub>The Company of Biologists Ltd</pub><pmid>29899136</pmid><doi>10.1242/dev.166215</doi><orcidid>https://orcid.org/0000-0003-3506-1287</orcidid><oa>free_for_read</oa></addata></record> |
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source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection; Company of Biologists |
subjects | Animals Body Patterning - genetics Cell Differentiation - genetics Cell Lineage - genetics Cells, Cultured Embryo, Mammalian Gene Expression Profiling Gene Expression Regulation, Developmental Human Development Humans Mesoderm - cytology Mesoderm - embryology Mesoderm - physiology Mice Neural Stem Cells - cytology Neural Stem Cells - physiology Neurogenesis - genetics Neurogenesis - physiology Spinal Cord - cytology Spinal Cord - physiology Stem Cells - cytology Stem Cells - physiology |
title | Neural differentiation, selection and transcriptomic profiling of human neuromesodermal progenitor-like cells in vitro |
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