Specification of murine ground state pluripotent stem cells to regional neuronal populations
Pluripotent stem cells (PSCs) are a valuable tool for interrogating development, disease modelling, drug discovery and transplantation. Despite the burgeoned capability to fate restrict human PSCs to specific neural lineages, comparative protocols for mouse PSCs have not similarly advanced. Mouse pr...
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| Veröffentlicht in: | Scientific reports 2017-11, Vol.7 (1), p.16001-16, Article 16001 |
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| creator | Alsanie, Walaa F. Niclis, Jonathan C. Hunt, Cameron P. De Luzy, Isabelle R. Penna, Vanessa Bye, Christopher R. Pouton, Colin W. Haynes, John Firas, Jaber Thompson, Lachlan H. Parish, Clare L. |
| description | Pluripotent stem cells (PSCs) are a valuable tool for interrogating development, disease modelling, drug discovery and transplantation. Despite the burgeoned capability to fate restrict human PSCs to specific neural lineages, comparative protocols for mouse PSCs have not similarly advanced. Mouse protocols fail to recapitulate neural development, consequently yielding highly heterogeneous populations, yet mouse PSCs remain a valuable scientific tool as differentiation is rapid, cost effective and an extensive repertoire of transgenic lines provides an invaluable resource for understanding biology. Here we developed protocols for neural fate restriction of mouse PSCs, using knowledge of embryonic development and recent progress with human equivalents. These methodologies rely upon naïve ground-state PSCs temporarily transitioning through LIF-responsive stage prior to neural induction and rapid exposure to regional morphogens. Neural subtypes generated included those of the dorsal forebrain, ventral forebrain, ventral midbrain and hindbrain. This rapid specification, without feeder layers or embryoid-body formation, resulted in high proportions of correctly specified progenitors and neurons with robust reproducibility. These generated neural progenitors/neurons will provide a valuable resource to further understand development, as well disorders affecting specific neuronal subpopulations. |
| doi_str_mv | 10.1038/s41598-017-16248-x |
| format | Article |
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Despite the burgeoned capability to fate restrict human PSCs to specific neural lineages, comparative protocols for mouse PSCs have not similarly advanced. Mouse protocols fail to recapitulate neural development, consequently yielding highly heterogeneous populations, yet mouse PSCs remain a valuable scientific tool as differentiation is rapid, cost effective and an extensive repertoire of transgenic lines provides an invaluable resource for understanding biology. Here we developed protocols for neural fate restriction of mouse PSCs, using knowledge of embryonic development and recent progress with human equivalents. These methodologies rely upon naïve ground-state PSCs temporarily transitioning through LIF-responsive stage prior to neural induction and rapid exposure to regional morphogens. Neural subtypes generated included those of the dorsal forebrain, ventral forebrain, ventral midbrain and hindbrain. This rapid specification, without feeder layers or embryoid-body formation, resulted in high proportions of correctly specified progenitors and neurons with robust reproducibility. These generated neural progenitors/neurons will provide a valuable resource to further understand development, as well disorders affecting specific neuronal subpopulations.</description><identifier>ISSN: 2045-2322</identifier><identifier>EISSN: 2045-2322</identifier><identifier>DOI: 10.1038/s41598-017-16248-x</identifier><identifier>PMID: 29167563</identifier><language>eng</language><publisher>London: Nature Publishing Group UK</publisher><subject>13 ; 13/100 ; 13/106 ; 13/31 ; 13/51 ; 631/378/2571 ; 631/532/1360 ; 631/532/2064 ; 631/532/2182 ; Animals ; Cell Differentiation - physiology ; Drug discovery ; Embryogenesis ; Embryonic growth stage ; Flow Cytometry ; Forebrain ; Hindbrain ; Humanities and Social Sciences ; Immunohistochemistry ; Induced Pluripotent Stem Cells - cytology ; Mesencephalon ; Mesencephalon - cytology ; Mice ; multidisciplinary ; Neural stem cells ; Neural Stem Cells - cytology ; Neural Stem Cells - metabolism ; Neurogenesis ; Neurogenesis - physiology ; Otx Transcription Factors - metabolism ; PAX6 Transcription Factor - metabolism ; Pluripotency ; Pluripotent Stem Cells - cytology ; Pluripotent Stem Cells - metabolism ; Prosencephalon - cytology ; Real-Time Polymerase Chain Reaction ; Rodents ; Science ; Science (multidisciplinary) ; Stem cell transplantation ; Stem cells ; Subpopulations ; Transplantation</subject><ispartof>Scientific reports, 2017-11, Vol.7 (1), p.16001-16, Article 16001</ispartof><rights>The Author(s) 2017</rights><rights>2017. 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These generated neural progenitors/neurons will provide a valuable resource to further understand development, as well disorders affecting specific neuronal subpopulations.</description><subject>13</subject><subject>13/100</subject><subject>13/106</subject><subject>13/31</subject><subject>13/51</subject><subject>631/378/2571</subject><subject>631/532/1360</subject><subject>631/532/2064</subject><subject>631/532/2182</subject><subject>Animals</subject><subject>Cell Differentiation - physiology</subject><subject>Drug discovery</subject><subject>Embryogenesis</subject><subject>Embryonic growth stage</subject><subject>Flow Cytometry</subject><subject>Forebrain</subject><subject>Hindbrain</subject><subject>Humanities and Social Sciences</subject><subject>Immunohistochemistry</subject><subject>Induced Pluripotent Stem Cells - cytology</subject><subject>Mesencephalon</subject><subject>Mesencephalon - cytology</subject><subject>Mice</subject><subject>multidisciplinary</subject><subject>Neural stem cells</subject><subject>Neural Stem Cells - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Scientific reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Alsanie, Walaa F.</au><au>Niclis, Jonathan C.</au><au>Hunt, Cameron P.</au><au>De Luzy, Isabelle R.</au><au>Penna, Vanessa</au><au>Bye, Christopher R.</au><au>Pouton, Colin W.</au><au>Haynes, John</au><au>Firas, Jaber</au><au>Thompson, Lachlan H.</au><au>Parish, Clare L.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Specification of murine ground state pluripotent stem cells to regional neuronal populations</atitle><jtitle>Scientific reports</jtitle><stitle>Sci Rep</stitle><addtitle>Sci Rep</addtitle><date>2017-11-22</date><risdate>2017</risdate><volume>7</volume><issue>1</issue><spage>16001</spage><epage>16</epage><pages>16001-16</pages><artnum>16001</artnum><issn>2045-2322</issn><eissn>2045-2322</eissn><abstract>Pluripotent stem cells (PSCs) are a valuable tool for interrogating development, disease modelling, drug discovery and transplantation. Despite the burgeoned capability to fate restrict human PSCs to specific neural lineages, comparative protocols for mouse PSCs have not similarly advanced. Mouse protocols fail to recapitulate neural development, consequently yielding highly heterogeneous populations, yet mouse PSCs remain a valuable scientific tool as differentiation is rapid, cost effective and an extensive repertoire of transgenic lines provides an invaluable resource for understanding biology. Here we developed protocols for neural fate restriction of mouse PSCs, using knowledge of embryonic development and recent progress with human equivalents. These methodologies rely upon naïve ground-state PSCs temporarily transitioning through LIF-responsive stage prior to neural induction and rapid exposure to regional morphogens. Neural subtypes generated included those of the dorsal forebrain, ventral forebrain, ventral midbrain and hindbrain. This rapid specification, without feeder layers or embryoid-body formation, resulted in high proportions of correctly specified progenitors and neurons with robust reproducibility. These generated neural progenitors/neurons will provide a valuable resource to further understand development, as well disorders affecting specific neuronal subpopulations.</abstract><cop>London</cop><pub>Nature Publishing Group UK</pub><pmid>29167563</pmid><doi>10.1038/s41598-017-16248-x</doi><tpages>16</tpages><orcidid>https://orcid.org/0000-0003-0224-3308</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | 13 13/100 13/106 13/31 13/51 631/378/2571 631/532/1360 631/532/2064 631/532/2182 Animals Cell Differentiation - physiology Drug discovery Embryogenesis Embryonic growth stage Flow Cytometry Forebrain Hindbrain Humanities and Social Sciences Immunohistochemistry Induced Pluripotent Stem Cells - cytology Mesencephalon Mesencephalon - cytology Mice multidisciplinary Neural stem cells Neural Stem Cells - cytology Neural Stem Cells - metabolism Neurogenesis Neurogenesis - physiology Otx Transcription Factors - metabolism PAX6 Transcription Factor - metabolism Pluripotency Pluripotent Stem Cells - cytology Pluripotent Stem Cells - metabolism Prosencephalon - cytology Real-Time Polymerase Chain Reaction Rodents Science Science (multidisciplinary) Stem cell transplantation Stem cells Subpopulations Transplantation |
| title | Specification of murine ground state pluripotent stem cells to regional neuronal populations |
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