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{"title"=>"Efficient Derivation of Multipotent Neural Stem/Progenitor Cells from Non-Human Primate Embryonic Stem Cells", "type"=>"journal", "authors"=>[{"first_name"=>"Hiroko", "last_name"=>"Shimada", "scopus_author_id"=>"35741769100"}, {"first_name"=>"Yohei", "last_name"=>"Okada", "scopus_author_id"=>"55322888400"}, {"first_name"=>"Keiji", "last_name"=>"Ibata", "scopus_author_id"=>"6603886557"}, {"first_name"=>"Hayao", "last_name"=>"Ebise", "scopus_author_id"=>"16635716100"}, {"first_name"=>"Shin ichi", "last_name"=>"Ota", "scopus_author_id"=>"35741265700"}, {"first_name"=>"Ikuo", "last_name"=>"Tomioka", "scopus_author_id"=>"6602817668"}, {"first_name"=>"Toshihiro", "last_name"=>"Nomura", "scopus_author_id"=>"57200800854"}, {"first_name"=>"Takuji", "last_name"=>"Maeda", "scopus_author_id"=>"26641444100"}, {"first_name"=>"Kazuhisa", "last_name"=>"Kohda", "scopus_author_id"=>"7102557490"}, {"first_name"=>"Michisuke", "last_name"=>"Yuzaki", "scopus_author_id"=>"7003817938"}, {"first_name"=>"Erika", "last_name"=>"Sasaki", "scopus_author_id"=>"55450501000"}, {"first_name"=>"Masaya", "last_name"=>"Nakamura", "scopus_author_id"=>"7405346352"}, {"first_name"=>"Hideyuki", "last_name"=>"Okano", "scopus_author_id"=>"7201660094"}], "year"=>2012, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "isbn"=>"1932-6203", "pui"=>"366053758", "sgr"=>"84869142111", "doi"=>"10.1371/journal.pone.0049469", "scopus"=>"2-s2.0-84869142111", "pmid"=>"23166679"}, "id"=>"16ff2ad1-cf30-371e-84e8-5cad22ddcb88", "abstract"=>"The common marmoset (Callithrix jacchus) is a small New World primate that has been used as a non-human primate model for various biomedical studies. We previously demonstrated that transplantation of neural stem/progenitor cells (NS/PCs) derived from mouse and human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) promote functional locomotor recovery of mouse spinal cord injury models. However, for the clinical application of such a therapeutic approach, we need to evaluate the efficacy and safety of pluripotent stem cell-derived NS/PCs not only by xenotransplantation, but also allotransplantation using non-human primate models to assess immunological rejection and tumorigenicity. In the present study, we established a culture method to efficiently derive NS/PCs as neurospheres from common marmoset ESCs. Marmoset ESC-derived neurospheres could be passaged repeatedly and showed sequential generation of neurons and astrocytes, similar to that of mouse ESC-derived NS/PCs, and gave rise to functional neurons as indicated by calcium imaging. Although marmoset ESC-derived NS/PCs could not differentiate into oligodendrocytes under default culture conditions, these cells could abundantly generate oligodendrocytes by incorporating additional signals that recapitulate in vivo neural development. Moreover, principal component analysis of microarray data demonstrated that marmoset ESC-derived NS/PCs acquired similar gene expression profiles to those of fetal brain-derived NS/PCs by repeated passaging. Therefore, marmoset ESC-derived NS/PCs may be useful not only for accurate evaluation by allotransplantation of NS/PCs into non-human primate models, but are also applicable to analysis of iPSCs established from transgenic disease model marmosets.", "link"=>"http://www.mendeley.com/research/efficient-derivation-multipotent-neural-stemprogenitor-cells-nonhuman-primate-embryonic-stem-cells", "reader_count"=>29, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>8, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>2, "Student > Master"=>6, "Other"=>1, "Lecturer"=>1, "Professor"=>3}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>8, "Student > Doctoral Student"=>1, "Student > Ph. D. Student"=>6, "Student > Postgraduate"=>2, "Student > Master"=>6, "Other"=>1, "Lecturer"=>1, "Professor"=>3}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>4, "Agricultural and Biological Sciences"=>15, "Medicine and Dentistry"=>6, "Neuroscience"=>4}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>6}, "Neuroscience"=>{"Neuroscience"=>4}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>15}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>4}}, "reader_count_by_country"=>{"United States"=>1, "Japan"=>2, "Chile"=>1, "Australia"=>1}, "group_count"=>0}

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• http://doi.org/10.1007/s00198-013-2566-4
• http://doi.org/10.1089/scd.2015.0244
• http://doi.org/10.1016/j.expneurol.2014.05.021
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• http://doi.org/10.1002/term.2072
• http://doi.org/10.3390/ijms19092788
• http://doi.org/10.1016/j.stemcr.2014.07.011
• http://doi.org/10.1038/npp.2016.41
• http://doi.org/10.1016/j.fob.2014.02.007
• http://doi.org/10.1002/adhm.201400042
• http://doi.org/10.5966/sctm.2014-0215
• http://doi.org/10.1038/s41536-019-0066-7
• http://doi.org/10.1016/j.neures.2019.09.005
• http://doi.org/10.1089/scd.2017.0069
• http://doi.org/10.1002/jbm.a.35466
• http://doi.org/10.1038/s41598-018-37990-w
• http://doi.org/10.1186/1756-6606-7-24
• http://doi.org/10.1016/j.neures.2014.04.002

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• {"id"=>"269439343033188352", "text"=>"#me NS/PCs from common marmoset ESCs: Multipotent neural stem/progenitor cells from non-human primate embryonic #sc's http://t.co/qC0gcrjt", "created_at"=>"2012-11-16T13:58:39Z", "user"=>"BiomedRevon", "user_name"=>"Mirko von Elstermann", "user_profile_image"=>"http://a0.twimg.com/profile_images/1518893315/Arcimboldo_Librarian_Stokholm_normal.jpg"}

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• {"files"=>["https://ndownloader.figshare.com/files/290944", "https://ndownloader.figshare.com/files/291009"], "description"=>"<div><p>The common marmoset (<em>Callithrix jacchus</em>) is a small New World primate that has been used as a non-human primate model for various biomedical studies. We previously demonstrated that transplantation of neural stem/progenitor cells (NS/PCs) derived from mouse and human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) promote functional locomotor recovery of mouse spinal cord injury models. However, for the clinical application of such a therapeutic approach, we need to evaluate the efficacy and safety of pluripotent stem cell-derived NS/PCs not only by xenotransplantation, but also allotransplantation using non-human primate models to assess immunological rejection and tumorigenicity. In the present study, we established a culture method to efficiently derive NS/PCs as neurospheres from common marmoset ESCs. Marmoset ESC-derived neurospheres could be passaged repeatedly and showed sequential generation of neurons and astrocytes, similar to that of mouse ESC-derived NS/PCs, and gave rise to functional neurons as indicated by calcium imaging. Although marmoset ESC-derived NS/PCs could not differentiate into oligodendrocytes under default culture conditions, these cells could abundantly generate oligodendrocytes by incorporating additional signals that recapitulate <em>in vivo</em> neural development. Moreover, principal component analysis of microarray data demonstrated that marmoset ESC-derived NS/PCs acquired similar gene expression profiles to those of fetal brain-derived NS/PCs by repeated passaging. Therefore, marmoset ESC-derived NS/PCs may be useful not only for accurate evaluation by allotransplantation of NS/PCs into non-human primate models, but are also applicable to analysis of iPSCs established from transgenic disease model marmosets.</p> </div>", "links"=>[], "tags"=>["derivation", "multipotent", "neural", "cells", "non-human", "primate", "embryonic", "cells"], "article_id"=>117278, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Hiroko Shimada", "Yohei Okada", "Keiji Ibata", "Hayao Ebise", "Shin-ichi Ota", "Ikuo Tomioka", "Toshihiro Nomura", "Takuji Maeda", "Kazuhisa Kohda", "Michisuke Yuzaki", "Erika Sasaki", "Masaya Nakamura", "Hideyuki Okano"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0049469.s001", "https://dx.doi.org/10.1371/journal.pone.0049469.s002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Efficient_Derivation_of_Multipotent_Neural_Stem_Progenitor_Cells_from_Non_Human_Primate_Embryonic_Stem_Cells__/117278", "title"=>"Efficient Derivation of Multipotent Neural Stem/Progenitor Cells from Non-Human Primate Embryonic Stem Cells", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2012-11-14 02:01:18"}
• {"files"=>["https://ndownloader.figshare.com/files/543633"], "description"=>"<p>(<b>A</b>) Protocol to derive neurospheres from marmoset ESCs by EB formation. EBs were cultured in suspension in ultra-low cluster dishes for 2 weeks in the presence of 3 µM dorsomorphin or 1×10⁻⁶ M RA. Dorsomorphin or RA were added on day 1 or 5 of EB formation, respectively. EBs were then dissociated and cultured in suspension for 12–14 days to form neurospheres in MHM medium containing 2% B27 and 20 ng/ml FGF-2. Primary neurospheres were dissociated and cultured in suspension again with FGF-2 to form secondary neurospheres. (<b>B</b>) Neurosphere formation rates are presented as the percentages of neurospheres among total cells plated. EBs treated with 3 µM dorsomorphin or 1×10⁻⁶ M RA were dissociated and cultured in MHM medium containing 2% B27 and 20 ng/ml FGF-2 at a density of 2.5×10⁴ cells/ml in an ultra-low cluster 96-well plate for 1 week, and then neurospheres larger than 50 µm in diameter were counted. Data are presented as the means ± SEM (<i>n</i> = 3). (<b>C</b>) Representative morphologies of EBs, primary neurospheres and secondary neurospheres under each condition. Scale bars, 100 µm for EBs, 200 µm for neurospheres.</p>", "links"=>[], "tags"=>["marmoset"], "article_id"=>214117, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Hiroko Shimada", "Yohei Okada", "Keiji Ibata", "Hayao Ebise", "Shin-ichi Ota", "Ikuo Tomioka", "Toshihiro Nomura", "Takuji Maeda", "Kazuhisa Kohda", "Michisuke Yuzaki", "Erika Sasaki", "Masaya Nakamura", "Hideyuki Okano"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0049469.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Neurosphere_formation_of_marmoset_ESCs_/214117", "title"=>"Neurosphere formation of marmoset ESCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-14 01:08:37"}
• {"files"=>["https://ndownloader.figshare.com/files/543764"], "description"=>"<p>(<b>A</b>) Marmoset ESC-derived primary and secondary neurospheres were dissociated and allowed to differentiate for 10 days, followed by immunocytochemical analysis of βIII-tubulin (neurons), GFAP (astrocytes), CNPase (oligodendrocytes) and Nestin (undifferentiated neural cells). Scale bars, 50 µm. (<b>B</b>) The proportions of cells positive for each cell type-specific marker are presented as the percentage of total cells counted by Hoechst 33258-stained nuclei. Data are presented as the means ± SEM (<i>n</i> = 3).</p>", "links"=>[], "tags"=>["potentials", "marmoset", "esc-derived"], "article_id"=>214242, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Hiroko Shimada", "Yohei Okada", "Keiji Ibata", "Hayao Ebise", "Shin-ichi Ota", "Ikuo Tomioka", "Toshihiro Nomura", "Takuji Maeda", "Kazuhisa Kohda", "Michisuke Yuzaki", "Erika Sasaki", "Masaya Nakamura", "Hideyuki Okano"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0049469.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Differentiation_potentials_of_marmoset_ESC_derived_NS_PCs_/214242", "title"=>"Differentiation potentials of marmoset ESC-derived NS/PCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-14 01:10:42"}
• {"files"=>["https://ndownloader.figshare.com/files/543902"], "description"=>"<p>qRT-PCR analysis of the expression of pluripotency markers (A: <i>Oct3/4</i> and <i>Nanog</i>), endodermal and mesodermal markers (B: <i>Afp</i> and <i>Gata4</i>, respectively), and neural markers (C: <i>Sox1</i> and <i>Pax6</i>) in ESCs, EBs and ESC-derived neurospheres. Fetal cell-derived (ganglionic eminence (GE), cortex and spinal cord) neurospheres and fetal tissues (heart and liver) were used as positive and negative controls, respectively. Data are presented as the means ± SEM (<i>n</i> = 3).</p>", "links"=>[], "tags"=>["type-specific", "markers", "differentiation"], "article_id"=>214379, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Hiroko Shimada", "Yohei Okada", "Keiji Ibata", "Hayao Ebise", "Shin-ichi Ota", "Ikuo Tomioka", "Toshihiro Nomura", "Takuji Maeda", "Kazuhisa Kohda", "Michisuke Yuzaki", "Erika Sasaki", "Masaya Nakamura", "Hideyuki Okano"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0049469.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Expression_of_cell_type_specific_markers_at_each_differentiation_step_/214379", "title"=>"Expression of cell type-specific markers at each differentiation step.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-14 01:12:59"}
• {"files"=>["https://ndownloader.figshare.com/files/543998"], "description"=>"<p>Rostrocaudal regional identities of marmoset ESC-derived neurospheres. qRT-PCR analysis of the expression of forebrain marker <i>Foxg1</i>, midbrain marker <i>Otx2</i> and spinal cord markers <i>Hoxc4</i> and <i>Hoxc6</i> in ESCs, EBs and ESC-derived neurospheres. Fetal cell-derived (ganglionic eminence (GE), cortex and spinal cord) neurospheres and fetal tissues (heart and liver) were used as positive and negative controls, respectively. Data are presented as the means ± SEM (<i>n</i> = 3).</p>", "links"=>[], "tags"=>["marmoset", "esc-derived"], "article_id"=>214487, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Hiroko Shimada", "Yohei Okada", "Keiji Ibata", "Hayao Ebise", "Shin-ichi Ota", "Ikuo Tomioka", "Toshihiro Nomura", "Takuji Maeda", "Kazuhisa Kohda", "Michisuke Yuzaki", "Erika Sasaki", "Masaya Nakamura", "Hideyuki Okano"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0049469.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Rostrocaudal_regulation_of_marmoset_ESC_derived_NS_PCs_/214487", "title"=>"Rostrocaudal regulation of marmoset ESC-derived NS/PCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-14 01:14:47"}
• {"files"=>["https://ndownloader.figshare.com/files/544129"], "description"=>"<p>(<b>A</b>) Protocol for derivation of NS/PCs from marmoset ESCs, which efficiently generate oligodendrocytes. EBs were cultured in the presence of 1×10⁻⁶ M RA and 2 µM purmorphamine for 2 weeks. RA and purmorphamine were added on day 5 and 7 of EB formation, respectively. EBs were then dissociated and cultured in suspension for 8–10 days to form neurospheres in MHM medium containing 2% B27, 20 ng/ml FGF-2, 20 ng/ml EGF, 1 µM purmorphamine and an oligodendrocyte cocktail. Primary neurospheres were dissociated and cultured under the same condition to form secondary neurospheres. (<b>B</b>) Differentiation of neurospheres derived from EBs treated with RA and purmorphamine. Primary and secondary neurospheres were dissociated and differentiated on poly-L-ornithine/laminin-coated coverslips at a density of 2×10⁴ cells/ml in MHM medium containing 2% B27 and an oligodendrocyte cocktail for 30–35 days, followed by immunocytochemical analysis of βIII-tubulin (neurons), GFAP (astrocytes) and O4 (oligodendrocytes). The obtained NS/PCs could efficiently give rise to oligodendrocytes. Scale bars, 100 µm. (<b>C</b>) The proportions of cells positive for each cell type-specific marker in differentiated secondary neurospheres, which could generate oligodendrocytes, are presented as the percentage of total cells counted by Hoechst 33258-stained nuclei. Data are presented as the means ± SEM (<i>n</i> = 3). (<b>D</b>) An enlarged image of βIII-tubulin-positive neurons, GFAP-positive astrocytes and O4-positive oligodendrocytes differentiated from secondary neurospheres. Scale bars, 100 µm. (<b>E</b>) Immunocytochemical analysis of secondary neurospheres for various markers of oligodendrocyte lineages, including PDGFR (OPCs), CNPase (oligodendrocytes) and MBP (mature oligodendrocytes). Scale bars, 50 µm.</p>", "links"=>[], "tags"=>["derivation", "oligodendrocytes", "marmoset"], "article_id"=>214612, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Hiroko Shimada", "Yohei Okada", "Keiji Ibata", "Hayao Ebise", "Shin-ichi Ota", "Ikuo Tomioka", "Toshihiro Nomura", "Takuji Maeda", "Kazuhisa Kohda", "Michisuke Yuzaki", "Erika Sasaki", "Masaya Nakamura", "Hideyuki Okano"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0049469.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Efficient_derivation_of_oligodendrocytes_from_marmoset_ESCs_/214612", "title"=>"Efficient derivation of oligodendrocytes from marmoset ESCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-14 01:16:52"}
• {"files"=>["https://ndownloader.figshare.com/files/544268"], "description"=>"<p>NS/PCs that could generate oligodendrocytes were allowed to differentiate for 10–11 weeks. Then, [Ca²⁺]_i in differentiated cells were measured by the intensity of fluo-4 fluorescence upon electrical field stimulation at 40 Hz for 5 seconds. Electrical field stimulation induced an elevation of [Ca²⁺]_i in several cells (<b>A</b>). The intensity of the increase of [Ca²⁺]_i was quantified (<b>B</b>). The increase of [Ca²⁺]_i was suppressed by TTX that specifically blocks voltage-gated sodium channels. Scale bars, 50 µm.</p>", "links"=>[], "tags"=>["esc-derived"], "article_id"=>214754, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Hiroko Shimada", "Yohei Okada", "Keiji Ibata", "Hayao Ebise", "Shin-ichi Ota", "Ikuo Tomioka", "Toshihiro Nomura", "Takuji Maeda", "Kazuhisa Kohda", "Michisuke Yuzaki", "Erika Sasaki", "Masaya Nakamura", "Hideyuki Okano"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0049469.g006"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Marmoset_ESC_derived_NS_PCs_give_rise_to_functional_neurons_/214754", "title"=>"Marmoset ESC-derived NS/PCs give rise to functional neurons.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-14 01:19:14"}
• {"files"=>["https://ndownloader.figshare.com/files/544369"], "description"=>"<p>PCA of all microarray data was conducted on cells at each differentiation step. Undifferentiated ESCs acquired gene expression profiles close to those of fetal neurospheres after passaging into secondary neurospheres.</p>", "links"=>[], "tags"=>["esc-derived", "neurospheres", "properties", "observed", "marmoset", "embryonic"], "article_id"=>214853, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Hiroko Shimada", "Yohei Okada", "Keiji Ibata", "Hayao Ebise", "Shin-ichi Ota", "Ikuo Tomioka", "Toshihiro Nomura", "Takuji Maeda", "Kazuhisa Kohda", "Michisuke Yuzaki", "Erika Sasaki", "Masaya Nakamura", "Hideyuki Okano"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0049469.g007"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Marmoset_ESC_derived_neurospheres_acquire_similar_properties_to_those_observed_in_marmoset_embryonic_neurospheres_/214853", "title"=>"Marmoset ESC-derived neurospheres acquire similar properties to those observed in marmoset embryonic neurospheres.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-14 01:20:53"}
• {"files"=>["https://ndownloader.figshare.com/files/544563"], "description"=>"<p>NS/PCs derived from EBs treated with both RA and purmorphamine, were transplanted into the right striatum of 6–8-week-old female NOD/SCID mice. 4 weeks after the injection, mice were sacrificed and processed for immunohistochemical analysis with the antibodies against markers for neurons (Hu), astrocytes (GFAP), oligodendrocytes (APC), and neural progenitors (Nestin). The marmoset ESC-derived NS/PCs could give rise to neurons, astrocytes and oligodendrocytes <i>in vivo</i>, as well. Scale bars, 20 µm.</p>", "links"=>[], "tags"=>["marmoset", "esc-derived"], "article_id"=>215048, "categories"=>["Neuroscience", "Developmental Biology"], "users"=>["Hiroko Shimada", "Yohei Okada", "Keiji Ibata", "Hayao Ebise", "Shin-ichi Ota", "Ikuo Tomioka", "Toshihiro Nomura", "Takuji Maeda", "Kazuhisa Kohda", "Michisuke Yuzaki", "Erika Sasaki", "Masaya Nakamura", "Hideyuki Okano"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0049469.g008"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Transplantation_of_marmoset_ESC_derived_neurospheres_/215048", "title"=>"Transplantation of marmoset ESC-derived neurospheres.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2012-11-14 01:24:08"}

PMC Usage Stats | Further Information

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Relative Metric

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F1000Prime