Dual Small-Molecule Targeting of SMAD Signaling Stimulates Human Induced Pluripotent Stem Cells toward Neural Lineages
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{"title"=>"Dual small-molecule targeting of SMAD signaling stimulates human induced pluripotent stem cells toward neural lineages", "type"=>"journal", "authors"=>[{"first_name"=>"Methichit", "last_name"=>"Wattanapanitch", "scopus_author_id"=>"35336151100"}, {"first_name"=>"Nuttha", "last_name"=>"Klincumhom", "scopus_author_id"=>"26642113000"}, {"first_name"=>"Porntip", "last_name"=>"Potirat", "scopus_author_id"=>"57173190500"}, {"first_name"=>"Rattaya", "last_name"=>"Amornpisutt", "scopus_author_id"=>"55232443700"}, {"first_name"=>"Chanchao", "last_name"=>"Lorthongpanich", "scopus_author_id"=>"8719782800"}, {"first_name"=>"Yaowalak", "last_name"=>"U-Pratya", "scopus_author_id"=>"6507352276"}, {"first_name"=>"Chuti", "last_name"=>"Laowtammathron", "scopus_author_id"=>"15832539200"}, {"first_name"=>"Pakpoom", "last_name"=>"Kheolamai", "scopus_author_id"=>"26537669600"}, {"first_name"=>"Niphon", "last_name"=>"Poungvarin", "scopus_author_id"=>"7005666706"}, {"first_name"=>"Surapol", "last_name"=>"Iaragrisil", "scopus_author_id"=>"57172928700"}], "year"=>2014, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "scopus"=>"2-s2.0-84946585569", "pui"=>"608994022", "doi"=>"10.1371/journal.pone.0106952", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "sgr"=>"84946585569", "pmid"=>"25207966"}, "id"=>"9f835ec3-f005-3a48-b18c-38d9ed094024", "abstract"=>"Incurable neurological disorders such as Parkinson's disease (PD), Huntington's disease (HD), and Alzheimer's disease (AD) are very common and can be life-threatening because of their progressive disease symptoms with limited treatment options. To provide an alternative renewable cell source for cell-based transplantation and as study models for neurological diseases, we generated induced pluripotent stem cells (iPSCs) from human dermal fibroblasts (HDFs) and then differentiated them into neural progenitor cells (NPCs) and mature neurons by dual SMAD signaling inhibitors. Reprogramming efficiency was improved by supplementing the histone deacethylase inhibitor, valproic acid (VPA), and inhibitor of p160-Rho associated coiled-coil kinase (ROCK), Y-27632, after retroviral transduction. We obtained a number of iPS colonies that shared similar characteristics with human embryonic stem cells in terms of their morphology, cell surface antigens, pluripotency-associated gene and protein expressions as well as their in vitro and in vivo differentiation potentials. After treatment with Noggin and SB431542, inhibitors of the SMAD signaling pathway, HDF-iPSCs demonstrated rapid and efficient differentiation into neural lineages. Six days after neural induction, neuroepithelial cells (NEPCs) were observed in the adherent monolayer culture, which had the ability to differentiate further into NPCs and neurons, as characterized by their morphology and the expression of neuron-specific transcripts and proteins. We propose that our study may be applied to generate neurological disease patient-specific iPSCs allowing better understanding of disease pathogenesis and drug sensitivity assays.", "link"=>"http://www.mendeley.com/research/dual-smallmolecule-targeting-smad-signaling-stimulates-human-induced-pluripotent-stem-cells-toward-n-2", "reader_count"=>51, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>2, "Student > Doctoral Student"=>4, "Researcher"=>11, "Student > Ph. D. Student"=>19, "Student > Master"=>8, "Other"=>2, "Student > Bachelor"=>4, "Professor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>2, "Student > Doctoral Student"=>4, "Researcher"=>11, "Student > Ph. D. Student"=>19, "Student > Master"=>8, "Other"=>2, "Student > Bachelor"=>4, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>3, "Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>14, "Agricultural and Biological Sciences"=>22, "Medicine and Dentistry"=>4, "Neuroscience"=>3, "Sports and Recreations"=>1, "Chemistry"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>3}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Neuroscience"=>{"Neuroscience"=>3}, "Chemistry"=>{"Chemistry"=>1}, "Sports and Recreations"=>{"Sports and Recreations"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>22}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>14}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"Sweden"=>1, "United Kingdom"=>2, "Germany"=>1}, "group_count"=>2}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1669236"], "description"=>"<p>(A) Schematic diagram represents a protocol to generate iPSCs from HDFs by retroviral transduction. (B) Morphology of HDFs on day 1 post-transduction (i), non ESC-like colonies on day 8 (ii), ESC-like colonies on day 13 (iii), iPS colonies on iHFF plate (iv) and an iPS colony on Matrigel-coated plate (v), scale bar (i) and (iii) = 200 µm, (ii), (iv) and (v) = 500 µm. (C) RT-qPCR analysis of pluripotency- and fibroblast-associated genes in HDF, HDF-iPSC1 and Chula2.hES cells. (D) RT-PCR analysis of pluripotent genes, OCT4 and NANOG, of all the established iPS clones, HDF-iPSC2-6, as compared to Chula2.hES cells. (E) Representative immunofluorescent images of pluripotent transcription factors and cell surface antigens of HDF-iPSC1 as compared to Chula2.hES cells, scale bar = 100 µm.</p>", "links"=>[], "tags"=>["iPS colonies", "Smad signaling", "invivo differentiation potentials", "study models", "disease pathogenesis", "histone deacethylase inhibitor", "protein expressions", "neuroepithelial cells", "npc", "disease symptoms", "hdf", "hd", "pd", "Neural Lineages Incurable", "monolayer culture", "NEPC", "treatment options", "Human induced pluripotent stem cells", "cell surface antigens", "sb", "valproic acid", "drug sensitivity assays", "cell source", "vpa", "progenitor cells", "Reprogramming efficiency"], "article_id"=>1166929, "categories"=>["Biological Sciences"], "users"=>["Methichit Wattanapanitch", "Nuttha Klincumhom", "Porntip Potirat", "Rattaya Amornpisutt", "Chanchao Lorthongpanich", "Yaowalak U-pratya", "Chuti Laowtammathron", "Pakpoom Kheolamai", "Niphon Poungvarin", "Surapol Issaragrisil"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106952.g001", "stats"=>{"downloads"=>4, "page_views"=>74, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Generation_and_characterization_of_iPSCs_from_human_dermal_fibroblasts_HDFs_/1166929", "title"=>"Generation and characterization of iPSCs from human dermal fibroblasts (HDFs).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-10 02:59:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/1669259", "https://ndownloader.figshare.com/files/1669260", "https://ndownloader.figshare.com/files/1669261", "https://ndownloader.figshare.com/files/1669262"], "description"=>"<div><p>Incurable neurological disorders such as Parkinson’s disease (PD), Huntington’s disease (HD), and Alzheimer’s disease (AD) are very common and can be life-threatening because of their progressive disease symptoms with limited treatment options. To provide an alternative renewable cell source for cell-based transplantation and as study models for neurological diseases, we generated induced pluripotent stem cells (iPSCs) from human dermal fibroblasts (HDFs) and then differentiated them into neural progenitor cells (NPCs) and mature neurons by dual SMAD signaling inhibitors. Reprogramming efficiency was improved by supplementing the histone deacethylase inhibitor, valproic acid (VPA), and inhibitor of p160-Rho associated coiled-coil kinase (ROCK), Y-27632, after retroviral transduction. We obtained a number of iPS colonies that shared similar characteristics with human embryonic stem cells in terms of their morphology, cell surface antigens, pluripotency-associated gene and protein expressions as well as their <i>in</i><i>vitro</i> and <i>in</i><i>vivo</i> differentiation potentials. After treatment with Noggin and SB431542, inhibitors of the SMAD signaling pathway, HDF-iPSCs demonstrated rapid and efficient differentiation into neural lineages. Six days after neural induction, neuroepithelial cells (NEPCs) were observed in the adherent monolayer culture, which had the ability to differentiate further into NPCs and neurons, as characterized by their morphology and the expression of neuron-specific transcripts and proteins. We propose that our study may be applied to generate neurological disease patient-specific iPSCs allowing better understanding of disease pathogenesis and drug sensitivity assays.</p></div>", "links"=>[], "tags"=>["iPS colonies", "Smad signaling", "invivo differentiation potentials", "study models", "disease pathogenesis", "histone deacethylase inhibitor", "protein expressions", "neuroepithelial cells", "npc", "disease symptoms", "hdf", "hd", "pd", "Neural Lineages Incurable", "monolayer culture", "NEPC", "treatment options", "Human induced pluripotent stem cells", "cell surface antigens", "sb", "valproic acid", "drug sensitivity assays", "cell source", "vpa", "progenitor cells", "Reprogramming efficiency"], "article_id"=>1166947, "categories"=>["Biological Sciences"], "users"=>["Methichit Wattanapanitch", "Nuttha Klincumhom", "Porntip Potirat", "Rattaya Amornpisutt", "Chanchao Lorthongpanich", "Yaowalak U-pratya", "Chuti Laowtammathron", "Pakpoom Kheolamai", "Niphon Poungvarin", "Surapol Issaragrisil"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0106952.s001", "https://dx.doi.org/10.1371/journal.pone.0106952.s002", "https://dx.doi.org/10.1371/journal.pone.0106952.s003", "https://dx.doi.org/10.1371/journal.pone.0106952.s004"], "stats"=>{"downloads"=>6, "page_views"=>13, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Dual_Small_Molecule_Targeting_of_SMAD_Signaling_Stimulates_Human_Induced_Pluripotent_Stem_Cells_toward_Neural_Lineages_/1166947", "title"=>"Dual Small-Molecule Targeting of SMAD Signaling Stimulates Human Induced Pluripotent Stem Cells toward Neural Lineages", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2014-09-10 02:59:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/1669240"], "description"=>"<p>(A) Representative micrographs show morphology of the embryoid body (EB) of HDF-iPSC1 cells on day 1 in suspension culture (i), differentiated EB turned into neurons (ii), adipocytes (iii) and beating cardiomyocytes (iv), scale bar (i) = 500 µm, (ii) and (iii) = 50 µm and (iv) = 200 µm. (B) RT-qPCR analysis of pluripotent and lineage markers of the differentiated EB generated from HDF-iPSC1 and Chula2.hES cells as compared to the undifferentiated HDF-iPSC1 and Chula2.hES cells. (C) Immunofluorescent analysis showed the expression of lineage markers, AFP (endoderm), SMA (mesoderm) and NESTIN (ectoderm) of the EB outgrowth generated from HDF-iPSC1 cells after 4 weeks of differentiation, scale bar (i–iv) = 50 µm, (v) and (vi) = 100 µm. (D) Hematoxylin and eosin (H&E) staining of teratomas derived from HDF-iPSC1 cells at 8 weeks post-implantation into nude mice. The teratomas contained heterogeneous tissues of all three embryonic germ layers including neural rosettes (ectoderm), cartilage (mesoderm) and glandular structure (endoderm). (E) Representative karyotype analysis of HDF-iPSC1 cells at passage 32 shows normal karyotype (46, XY).</p>", "links"=>[], "tags"=>["iPS colonies", "Smad signaling", "invivo differentiation potentials", "study models", "disease pathogenesis", "histone deacethylase inhibitor", "protein expressions", "neuroepithelial cells", "npc", "disease symptoms", "hdf", "hd", "pd", "Neural Lineages Incurable", "monolayer culture", "NEPC", "treatment options", "Human induced pluripotent stem cells", "cell surface antigens", "sb", "valproic acid", "drug sensitivity assays", "cell source", "vpa", "progenitor cells", "Reprogramming efficiency"], "article_id"=>1166933, "categories"=>["Biological Sciences"], "users"=>["Methichit Wattanapanitch", "Nuttha Klincumhom", "Porntip Potirat", "Rattaya Amornpisutt", "Chanchao Lorthongpanich", "Yaowalak U-pratya", "Chuti Laowtammathron", "Pakpoom Kheolamai", "Niphon Poungvarin", "Surapol Issaragrisil"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106952.g002", "stats"=>{"downloads"=>0, "page_views"=>28, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_In_vitro_and_in_vivo_differentiation_and_karyotypic_analysis_of_HDF_iPSC1_cells_/1166933", "title"=>"<i>In vitro</i> and <i>in</i><i>vivo</i> differentiation, and karyotypic analysis of HDF-iPSC1 cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-10 02:59:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/1669244"], "description"=>"<p>(A) Schematic diagram represents the strategy to differentiate HDF-derived iPSCs into neural lineages using SMAD signaling inhibitors. (B) Morphological changes in HDF-iPSC1 cultures during neural differentiation; undifferentiated HDF-iPSC1 cells on day -1 (i), differentiated cells on day 2 (ii) and day 6 (iii), representative immunofluorescent images show the NEPC on day 6 (iv), NPC on day 9 (v) and post-mitotic neuron on day 28 (vi) of differentiation as examined by PAX6, NESTIN and TUJ1 markers, respectively. Cell nuclei were stained with DAPI. Scale bar = 100 µm. (C) RT-qPCR analysis of pluripotency- and neural-associated genes in NEPCs on day 6 and mature neuron on day 28 of differentiation as compared to those of undifferentiated HDF-iPSC1 cells (arbitrarily set at 1).</p>", "links"=>[], "tags"=>["iPS colonies", "Smad signaling", "invivo differentiation potentials", "study models", "disease pathogenesis", "histone deacethylase inhibitor", "protein expressions", "neuroepithelial cells", "npc", "disease symptoms", "hdf", "hd", "pd", "Neural Lineages Incurable", "monolayer culture", "NEPC", "treatment options", "Human induced pluripotent stem cells", "cell surface antigens", "sb", "valproic acid", "drug sensitivity assays", "cell source", "vpa", "progenitor cells", "Reprogramming efficiency"], "article_id"=>1166937, "categories"=>["Biological Sciences"], "users"=>["Methichit Wattanapanitch", "Nuttha Klincumhom", "Porntip Potirat", "Rattaya Amornpisutt", "Chanchao Lorthongpanich", "Yaowalak U-pratya", "Chuti Laowtammathron", "Pakpoom Kheolamai", "Niphon Poungvarin", "Surapol Issaragrisil"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0106952.g003", "stats"=>{"downloads"=>5, "page_views"=>96, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Neural_differentiation_of_HDF_derived_iPSCs_and_characterization_of_differentiated_neurons_/1166937", "title"=>"Neural differentiation of HDF-derived iPSCs and characterization of differentiated neurons.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2014-09-10 02:59:55"}

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Relative Metric

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