Generation of a defined and uniform population of CNS progenitors and neurons from mouse embryonic stem cells
A detailed protocol is described allowing the generation of essentially pure populations of glutamatergic neurons from mouse embryonic stem (ES) cells. It is based on the culture of ES cells that are kept undifferentiated by repeated splitting and subsequently amplified as non-adherent cell aggregat...
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| Veröffentlicht in: | Nature protocols 2007-05, Vol.2 (5), p.1034-1043 |
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| creator | Bibel, Miriam Richter, Jens Lacroix, Emmanuel Barde, Yves-Alain |
| description | A detailed protocol is described allowing the generation of essentially pure populations of glutamatergic neurons from mouse embryonic stem (ES) cells. It is based on the culture of ES cells that are kept undifferentiated by repeated splitting and subsequently amplified as non-adherent cell aggregates. Treatment with retinoic acid causes these ES cells to essentially become neural progenitors with the characteristics of Pax6-positive radial glial cells. As they do
in vivo
, these progenitors differentiate in glutamatergic pyramidal neurons that form functional synaptic contacts and can be kept in culture for long periods of time. This protocol does not require the use of ES lines expressing resistance or fluorescent markers and can thus be applied in principle to any wild-type or mutant ES line of interest. At least 2 weeks are required from starting ES cell culture until plating progenitors and differentiating neurons establish synaptic transmission within about 10 days. |
| doi_str_mv | 10.1038/nprot.2007.147 |
| format | Article |
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in vivo
, these progenitors differentiate in glutamatergic pyramidal neurons that form functional synaptic contacts and can be kept in culture for long periods of time. This protocol does not require the use of ES lines expressing resistance or fluorescent markers and can thus be applied in principle to any wild-type or mutant ES line of interest. At least 2 weeks are required from starting ES cell culture until plating progenitors and differentiating neurons establish synaptic transmission within about 10 days.</description><identifier>ISSN: 1754-2189</identifier><identifier>EISSN: 1750-2799</identifier><identifier>DOI: 10.1038/nprot.2007.147</identifier><identifier>PMID: 17546008</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>Aggregates ; Analytical Chemistry ; Animals ; Biological Techniques ; Biomedical and Life Sciences ; Cell culture ; Cell Culture Techniques - methods ; Cell cycle ; Cell differentiation ; Cell Differentiation - drug effects ; Central nervous system ; Central Nervous System - cytology ; Computational Biology/Bioinformatics ; Embryonic stem cells ; Embryonic Stem Cells - cytology ; Embryos ; Growth factors ; Life Sciences ; Mice ; Microarrays ; Mutation ; Nervous system ; Neurons - cytology ; Neurosciences ; Organic Chemistry ; Physiological aspects ; protocol ; Spinal cord ; Stem cells ; Tretinoin - pharmacology</subject><ispartof>Nature protocols, 2007-05, Vol.2 (5), p.1034-1043</ispartof><rights>Springer Nature Limited 2007</rights><rights>COPYRIGHT 2007 Nature Publishing Group</rights><rights>Copyright Nature Publishing Group May 2007</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c664t-eb5e40e5fa49d0b0d85760e4f7de1cbd7c7555f8e880bac6744ec7f362a363a23</citedby><cites>FETCH-LOGICAL-c664t-eb5e40e5fa49d0b0d85760e4f7de1cbd7c7555f8e880bac6744ec7f362a363a23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1038/nprot.2007.147$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1038/nprot.2007.147$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27915,27916,41479,42548,51310</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17546008$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Bibel, Miriam</creatorcontrib><creatorcontrib>Richter, Jens</creatorcontrib><creatorcontrib>Lacroix, Emmanuel</creatorcontrib><creatorcontrib>Barde, Yves-Alain</creatorcontrib><title>Generation of a defined and uniform population of CNS progenitors and neurons from mouse embryonic stem cells</title><title>Nature protocols</title><addtitle>Nat Protoc</addtitle><addtitle>Nat Protoc</addtitle><description>A detailed protocol is described allowing the generation of essentially pure populations of glutamatergic neurons from mouse embryonic stem (ES) cells. It is based on the culture of ES cells that are kept undifferentiated by repeated splitting and subsequently amplified as non-adherent cell aggregates. Treatment with retinoic acid causes these ES cells to essentially become neural progenitors with the characteristics of Pax6-positive radial glial cells. As they do
in vivo
, these progenitors differentiate in glutamatergic pyramidal neurons that form functional synaptic contacts and can be kept in culture for long periods of time. This protocol does not require the use of ES lines expressing resistance or fluorescent markers and can thus be applied in principle to any wild-type or mutant ES line of interest. 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Academic</collection><jtitle>Nature protocols</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Bibel, Miriam</au><au>Richter, Jens</au><au>Lacroix, Emmanuel</au><au>Barde, Yves-Alain</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Generation of a defined and uniform population of CNS progenitors and neurons from mouse embryonic stem cells</atitle><jtitle>Nature protocols</jtitle><stitle>Nat Protoc</stitle><addtitle>Nat Protoc</addtitle><date>2007-05-01</date><risdate>2007</risdate><volume>2</volume><issue>5</issue><spage>1034</spage><epage>1043</epage><pages>1034-1043</pages><issn>1754-2189</issn><eissn>1750-2799</eissn><abstract>A detailed protocol is described allowing the generation of essentially pure populations of glutamatergic neurons from mouse embryonic stem (ES) cells. It is based on the culture of ES cells that are kept undifferentiated by repeated splitting and subsequently amplified as non-adherent cell aggregates. Treatment with retinoic acid causes these ES cells to essentially become neural progenitors with the characteristics of Pax6-positive radial glial cells. As they do
in vivo
, these progenitors differentiate in glutamatergic pyramidal neurons that form functional synaptic contacts and can be kept in culture for long periods of time. This protocol does not require the use of ES lines expressing resistance or fluorescent markers and can thus be applied in principle to any wild-type or mutant ES line of interest. At least 2 weeks are required from starting ES cell culture until plating progenitors and differentiating neurons establish synaptic transmission within about 10 days.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>17546008</pmid><doi>10.1038/nprot.2007.147</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Aggregates Analytical Chemistry Animals Biological Techniques Biomedical and Life Sciences Cell culture Cell Culture Techniques - methods Cell cycle Cell differentiation Cell Differentiation - drug effects Central nervous system Central Nervous System - cytology Computational Biology/Bioinformatics Embryonic stem cells Embryonic Stem Cells - cytology Embryos Growth factors Life Sciences Mice Microarrays Mutation Nervous system Neurons - cytology Neurosciences Organic Chemistry Physiological aspects protocol Spinal cord Stem cells Tretinoin - pharmacology |
| title | Generation of a defined and uniform population of CNS progenitors and neurons from mouse embryonic stem cells |
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