Glycogen Synthase Kinase 3 (GSK3) Inhibitor, SB-216763, Promotes Pluripotency in Mouse Embryonic Stem Cells
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{"title"=>"Glycogen Synthase Kinase 3 (GSK3) inhibitor, SB-216763, promotes pluripotency in mouse embryonic stem cells", "type"=>"journal", "authors"=>[{"first_name"=>"Leslie A.", "last_name"=>"Kirby", "scopus_author_id"=>"55259640300"}, {"first_name"=>"Jason T.", "last_name"=>"Schott", "scopus_author_id"=>"57197885435"}, {"first_name"=>"Brenda L.", "last_name"=>"Noble", "scopus_author_id"=>"55260266900"}, {"first_name"=>"Daniel C.", "last_name"=>"Mendez", "scopus_author_id"=>"57196681358"}, {"first_name"=>"Paul S.", "last_name"=>"Caseley", "scopus_author_id"=>"55260412400"}, {"first_name"=>"Sarah C.", "last_name"=>"Peterson", "scopus_author_id"=>"55260252400"}, {"first_name"=>"Tyler J.", "last_name"=>"Routledge", "scopus_author_id"=>"55259900700"}, {"first_name"=>"Nilay V.", "last_name"=>"Patel", "scopus_author_id"=>"57198804099"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"pmid"=>"22745733", "doi"=>"10.1371/journal.pone.0039329", "sgr"=>"84862746199", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "scopus"=>"2-s2.0-84862746199", "issn"=>"19326203", "pui"=>"365093325"}, "id"=>"fa7a4d95-674b-3293-8736-15474083f5c1", "abstract"=>"Canonical Wnt/β-catenin signaling has been suggested to promote self-renewal of pluripotent mouse and human embryonic stem cells. Here, we show that SB-216763, a glycogen synthase kinase-3 (GSK3) inhibitor, can maintain mouse embryonic stem cells (mESCs) in a pluripotent state in the absence of exogenous leukemia inhibitory factor (LIF) when cultured on mouse embryonic fibroblasts (MEFs). MESCs maintained with SB-216763 for one month were morphologically indistinguishable from LIF-treated mESCs and expressed pluripotent-specific genes Oct4, Sox2, and Nanog. Furthermore, Nanog immunostaining was more homogenous in SB-216763-treated colonies compared to LIF. Embryoid bodies (EBs) prepared from these mESCs expressed early-stage markers for all three germ layers, and could efficiently differentiate into cardiac-like cells and MAP2-immunoreactive neurons. To our knowledge, SB-216763 is the first GSK3 inhibitor that can promote self-renewal of mESC co-cultured with MEFs for more than two months.", "link"=>"http://www.mendeley.com/research/glycogen-synthase-kinase-3-gsk3-inhibitor-sb216763-promotes-pluripotency-mouse-embryonic-stem-cells", "reader_count"=>72, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>5, "Researcher"=>13, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>25, "Student > Postgraduate"=>3, "Student > Master"=>7, "Other"=>4, "Student > Bachelor"=>11, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>5, "Researcher"=>13, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>25, "Student > Postgraduate"=>3, "Student > Master"=>7, "Other"=>4, "Student > Bachelor"=>11, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Engineering"=>1, "Biochemistry, Genetics and Molecular Biology"=>8, "Medicine and Dentistry"=>7, "Agricultural and Biological Sciences"=>43, "Neuroscience"=>2, "Sports and Recreations"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1, "Chemistry"=>4, "Psychology"=>1, "Social Sciences"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>7}, "Neuroscience"=>{"Neuroscience"=>2}, "Chemistry"=>{"Chemistry"=>4}, "Social Sciences"=>{"Social Sciences"=>1}, "Sports and Recreations"=>{"Sports and Recreations"=>1}, "Psychology"=>{"Psychology"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>43}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>8}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"United States"=>2, "Japan"=>1, "United Kingdom"=>3, "France"=>1, "Chile"=>1, "Australia"=>1, "Portugal"=>2, "Germany"=>1}, "group_count"=>6}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/618613"], "description"=>"<p>Cultures were maintained continuously with various doses of SB-216763 or LIF for 1 month and then immunostained for Oct–4. SB-216763 precipitated at doses ≥20 µM, and some of these particulates are visible in the phase-contrast images as black spots that fluoresced intensely green when excited at ∼480 nm. Scale bars represent 100 µm.</p>", "links"=>[], "tags"=>["mescs", "maintained", "sb-216763", "pluripotency"], "article_id"=>289101, "categories"=>["Neuroscience", "Genetics", "Developmental Biology"], "users"=>["Leslie A. Kirby", "Jason T. Schott", "Brenda L. Noble", "Daniel C. Mendez", "Paul S. Caseley", "Sarah C. Peterson", "Tyler J. Routledge", "Nilay V. Patel"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039329.g004", "stats"=>{"downloads"=>1, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_J1_mESCs_maintained_with_SB_216763_for_one_month_express_the_pluripotency_marker_Oct_8211_4_/289101", "title"=>"J1 mESCs maintained with SB-216763 for one month express the pluripotency marker Oct–4.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:31:41"}
  • {"files"=>["https://ndownloader.figshare.com/files/618425"], "description"=>"<p>The graph also shows expression of these markers in EBs derived from month-old LIF and SB-216763 mESCs. SYBR Green qPCR results for these four genes were normalized to β-actin housekeeping gene using qbasePLUS software. SB-216763 treated mESCs expressed Lrh-1 and Sox2 about 3-fold higher than LIF treated mESCs. Nanog expression was also moderately higher in the SB-216763-treated mESCs, whereas Oct-4 expression was nearly 50% higher in LIF-treated mESC. Expression of these genes in differentiated cells was generally lower than the mESCs maintained with either LIF or 10 µM SB-216763. The data are average ± S.D. of technical replicates from one of the two experiments.</p>", "links"=>[], "tags"=>["mescs", "maintained", "10", "sb-216763", "lif", "sox2"], "article_id"=>288908, "categories"=>["Neuroscience", "Genetics", "Developmental Biology"], "users"=>["Leslie A. Kirby", "Jason T. Schott", "Brenda L. Noble", "Daniel C. Mendez", "Paul S. Caseley", "Sarah C. Peterson", "Tyler J. Routledge", "Nilay V. Patel"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039329.g003", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_J1_mESCs_maintained_with_10_181_M_SB_216763_or_LIF_for_more_than_a_month_express_Oct4_Nanog_Sox2_and_Lrh_1_/288908", "title"=>"J1 mESCs maintained with 10 µM SB-216763 or LIF for more than a month express Oct4, Nanog, Sox2 and Lrh-1.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:28:28"}
  • {"files"=>["https://ndownloader.figshare.com/files/618327"], "description"=>"<p>(A) Phase-contrast images of mESCs treated with LIF (1,000 U/mL), or 10, 15, and 20 µM SB-216763 for more than a month. All doses of SB-216763 maintained mESCs as compact, elliptical or circular colonies characteristic of pluripotent cells. Morphology for mESC colonies maintained with SB-216763 was comparable to those in LIF supplemented medium. Scale bars represent 100 µm. (B) Twenty thousand mESCs from each treatment were seeded in a 6-well chamber at each passage and monitored for the percentage of pluripotent-like colonies two days after each passage. Data are expressed as the mean of five fields analyzed ± SEM. The percentage of pluripotent-like colonies in vehicle-treated cultures was significantly different from 15 µM and 20 µM SB-216763 samples at passage 4 (p<0.05) and from all treated samples from passage 5 onwards (p<0.05). (C) The average number of the total colonies and total pluripotent-like colonies observed between passages 4–7. The number of pluripotent-like colonies in vehicle-treated cultures was significantly lower than that for all other treatments (p<0.05). None of the SB-216763-treated samples were significantly different from LIF-treated mESCs for either the total number of colonies or the total number of pluripotent-like colonies.</p>", "links"=>[], "tags"=>["mescs", "maintained", "sb-216763", "pluripotent-like"], "article_id"=>288809, "categories"=>["Neuroscience", "Genetics", "Developmental Biology"], "users"=>["Leslie A. Kirby", "Jason T. Schott", "Brenda L. Noble", "Daniel C. Mendez", "Paul S. Caseley", "Sarah C. Peterson", "Tyler J. Routledge", "Nilay V. Patel"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039329.g002", "stats"=>{"downloads"=>2, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_J1_mESCs_maintained_with_SB_216763_have_a_pluripotent_like_morphology_/288809", "title"=>"J1 mESCs maintained with SB-216763 have a pluripotent-like morphology.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:26:49"}
  • {"files"=>["https://ndownloader.figshare.com/files/619203"], "description"=>"<p>(A) GFP fluorescence in representative colonies from each treatment. (B) Mean pluripotency rating of two independent experiments ± SEM. Forty colonies were rated from each condition for the intensity and uniformity of GFP fluorescence after a month in culture. Statistical analysis revealed that the cultures maintained with LIF, 3 µM and 10 µM SB-216763 were significantly different than mESCs maintained without LIF.</p>", "links"=>[], "tags"=>["sb-216763", "maintains", "millitrace", "nanog", "gfp", "mescs", "pluripotent-like"], "article_id"=>289685, "categories"=>["Neuroscience", "Genetics", "Developmental Biology"], "users"=>["Leslie A. Kirby", "Jason T. Schott", "Brenda L. Noble", "Daniel C. Mendez", "Paul S. Caseley", "Sarah C. Peterson", "Tyler J. Routledge", "Nilay V. Patel"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039329.g007", "stats"=>{"downloads"=>1, "page_views"=>120, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_10_181_M_SB_216763_maintains_MilliTrace_Nanog_GFP_Reporter_mESCs_in_a_pluripotent_like_state_for_more_than_a_month_/289685", "title"=>"10 µM SB-216763 maintains MilliTrace Nanog GFP Reporter mESCs in a pluripotent-like state for more than a month.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:41:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/618135"], "description"=>"<p>HEK293 cells (top panel) and J1 mESCs (bottom panel) were electroporated with the Super TOPFlash and pGL4.73 <i>Renilla</i> luciferase reporters. The cells were treated in quadruplicate wells with a vehicle control (0.1% DMSO) or the indicated doses of LIF, SB-216763, BIO, or CHIR-99021 for 24 hours. Cell lysates were then assayed for dual luciferase reporter activity. Firefly luciferase activity was normalized to <i>Renilla</i> luciferase activity for each sample, and then all results were normalized to the vehicle control. Data are expressed as the mean ± SEM from three independent trials.</p>", "links"=>[], "tags"=>["bio", "chir-99021", "induce"], "article_id"=>288622, "categories"=>["Neuroscience", "Genetics", "Developmental Biology"], "users"=>["Leslie A. Kirby", "Jason T. Schott", "Brenda L. Noble", "Daniel C. Mendez", "Paul S. Caseley", "Sarah C. Peterson", "Tyler J. Routledge", "Nilay V. Patel"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039329.g001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SB_216763_BIO_and_CHIR_99021_induce_expression_of_a_946_catenin_reporter_gene_in_a_dose_dependent_manner_/288622", "title"=>"SB-216763, BIO and CHIR-99021 induce expression of a β-catenin reporter gene in a dose dependent manner.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:23:42"}
  • {"files"=>["https://ndownloader.figshare.com/files/618780"], "description"=>"<p>MESCs maintained with either LIF or 10 µM SB-216763 for more than a month yielded pluripotent-like colonies. Immunostaining revealed that Nanog expression (green fluorescence) was homogeneous in mESC colonies maintained with SB-216763, whereas LIF-treated mESC colonies often displayed incomplete Nanog staining. Three representative colonies from each treatment are shown. Scale bars represent 40 µm.</p>", "links"=>[], "tags"=>["nanog", "pluripotency", "homogeneous", "j1", "mescs", "maintained", "10"], "article_id"=>289258, "categories"=>["Neuroscience", "Genetics", "Developmental Biology"], "users"=>["Leslie A. Kirby", "Jason T. Schott", "Brenda L. Noble", "Daniel C. Mendez", "Paul S. Caseley", "Sarah C. Peterson", "Tyler J. Routledge", "Nilay V. Patel"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039329.g005", "stats"=>{"downloads"=>1, "page_views"=>37, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_Nanog_pluripotency_marker_is_more_homogeneous_in_J1_mESCs_maintained_with_10_181_M_SB_216763_/289258", "title"=>"Expression of Nanog pluripotency marker is more homogeneous in J1 mESCs maintained with 10 µM SB-216763.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:34:18"}
  • {"files"=>["https://ndownloader.figshare.com/files/321601"], "description"=>"<div><p>Canonical Wnt/β-catenin signaling has been suggested to promote self-renewal of pluripotent mouse and human embryonic stem cells. Here, we show that SB-216763, a glycogen synthase kinase-3 (GSK3) inhibitor, can maintain mouse embryonic stem cells (mESCs) in a pluripotent state in the absence of exogenous leukemia inhibitory factor (LIF) when cultured on mouse embryonic fibroblasts (MEFs). MESCs maintained with SB-216763 for one month were morphologically indistinguishable from LIF-treated mESCs and expressed pluripotent-specific genes Oct4, Sox2, and Nanog. Furthermore, Nanog immunostaining was more homogenous in SB-216763-treated colonies compared to LIF. Embryoid bodies (EBs) prepared from these mESCs expressed early-stage markers for all three germ layers, and could efficiently differentiate into cardiac-like cells and MAP2-immunoreactive neurons. To our knowledge, SB-216763 is the first GSK3 inhibitor that can promote self-renewal of mESC co-cultured with MEFs for more than two months.</p> </div>", "links"=>[], "tags"=>["glycogen", "synthase", "kinase", "promotes", "pluripotency", "embryonic", "cells"], "article_id"=>123479, "categories"=>["Neuroscience", "Genetics", "Developmental Biology"], "users"=>["Leslie A. Kirby", "Jason T. Schott", "Brenda L. Noble", "Daniel C. Mendez", "Paul S. Caseley", "Sarah C. Peterson", "Tyler J. Routledge", "Nilay V. Patel"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039329", "stats"=>{"downloads"=>4, "page_views"=>10, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Glycogen_Synthase_Kinase_3_GSK3_Inhibitor_SB_216763_Promotes_Pluripotency_in_Mouse_Embryonic_Stem_Cells/123479", "title"=>"Glycogen Synthase Kinase 3 (GSK3) Inhibitor, SB-216763, Promotes Pluripotency in Mouse Embryonic Stem Cells", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 00:57:59"}
  • {"files"=>["https://ndownloader.figshare.com/files/618968"], "description"=>"<p>J1 mESCs co-cultured with MEFs were maintained continuously with vehicle (0.1% DMSO), 1,000 U/ml LIF, 10 or 20 µM SB-216763, 2 µM BIO, or 3 µM CHIR-99021. The cultures were passaged in parallel and 20,000–50,000 mESCs were plated on new MEF-covered 6-well plates every 3 days. A duplicate plate was stained for alkaline phosphatase (AP) activity at days 10, 20, and 30. Shown are representative images from one of two independent trials. Day 60 images were obtained from an independent culture maintained with vehicle (0.1% DMSO), 1,000 U/mL LIF, or 10 µM SB-216763. AP staining on day 60 was carried out on samples from a single trial. The scale bar shown in the day 10 vehicle image represents 100 µm and all images are presented at the same scale.</p>", "links"=>[], "tags"=>["bio", "j1", "mescs", "pluripotent"], "article_id"=>289452, "categories"=>["Neuroscience", "Genetics", "Developmental Biology"], "users"=>["Leslie A. Kirby", "Jason T. Schott", "Brenda L. Noble", "Daniel C. Mendez", "Paul S. Caseley", "Sarah C. Peterson", "Tyler J. Routledge", "Nilay V. Patel"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039329.g006", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_SB_216763_unlike_BIO_and_CHIR_99021_can_maintain_J1_mESCs_in_a_pluripotent_state_for_at_least_two_months_/289452", "title"=>"SB-216763, unlike BIO and CHIR-99021, can maintain J1 mESCs in a pluripotent state for at least two months.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:37:32"}
  • {"files"=>["https://ndownloader.figshare.com/files/619347"], "description"=>"<p>Cultures maintained with 10 µM SB-216763 or LIF for more than one month were differentiated into (A) cardiac-like cells, (B) neurons, and EBs expressing transcript markers for (C) ectoderm or neuroectoderm, (D) mesoderm, and (E) endoderm. (A) The number of flattened autonomously contracting EBs was assessed every other day after the 4-day old EBs were transferred to a 24-well chamber (one EB/well). More than 25% of the 20 EBs analyzed per condition showed some level of autonomous contraction. (B) Neurons were stained for the neuronal marker gene MAP-2 by immunocytochemistry 16 days after EB formation. Scale bars represent 40 µm. (C-E) EBs prepared from mESCs maintained with either 10 µM SB-216763 or LIF conditions expressed early-stage transcript markers for all three germ layers. Data are representative from one experiment ± SD.</p>", "links"=>[], "tags"=>["mescs", "maintained", "sb-216763", "differentiate", "germ"], "article_id"=>289833, "categories"=>["Neuroscience", "Genetics", "Developmental Biology"], "users"=>["Leslie A. Kirby", "Jason T. Schott", "Brenda L. Noble", "Daniel C. Mendez", "Paul S. Caseley", "Sarah C. Peterson", "Tyler J. Routledge", "Nilay V. Patel"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0039329.g008", "stats"=>{"downloads"=>3, "page_views"=>20, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_J1_mESCs_maintained_with_SB_216763_retain_their_ability_to_differentiate_into_all_three_germ_layers_/289833", "title"=>"J1 mESCs maintained with SB-216763 retain their ability to differentiate into all three germ layers.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-06-26 02:43:53"}

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Relative Metric

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