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{"title"=>"Generation of Germline-Competent rat induced pluripotent stem cells", "type"=>"journal", "authors"=>[{"first_name"=>"Sanae", "last_name"=>"Hamanaka", "scopus_author_id"=>"36899898200"}, {"first_name"=>"Tomoyuki", "last_name"=>"Yamaguchi", "scopus_author_id"=>"55694091300"}, {"first_name"=>"Toshihiro", "last_name"=>"Kobayashi", "scopus_author_id"=>"7406710741"}, {"first_name"=>"Megumi", "last_name"=>"Kato-Itoh", "scopus_author_id"=>"36673173900"}, {"first_name"=>"Satoshi", "last_name"=>"Yamazaki", "scopus_author_id"=>"35401033700"}, {"first_name"=>"Hideyuki", "last_name"=>"Sato", "scopus_author_id"=>"55316870800"}, {"first_name"=>"Ayumi", "last_name"=>"Umino", "scopus_author_id"=>"47761786700"}, {"first_name"=>"Yukiko", "last_name"=>"Wakiyama", "scopus_author_id"=>"35077976600"}, {"first_name"=>"Mami", "last_name"=>"Arai", "scopus_author_id"=>"56645076600"}, {"first_name"=>"Makoto", "last_name"=>"Sanbo", "scopus_author_id"=>"6507798871"}, {"first_name"=>"Masumi", "last_name"=>"Hirabayashi", "scopus_author_id"=>"35466283700"}, {"first_name"=>"Hiromitsu", "last_name"=>"Nakauchi", "scopus_author_id"=>"35393672800"}], "year"=>2011, "source"=>"PLoS ONE", "identifiers"=>{"pui"=>"362137002", "sgr"=>"79960330734", "issn"=>"19326203", "pmid"=>"21789202", "scopus"=>"2-s2.0-79960330734", "doi"=>"10.1371/journal.pone.0022008", "isbn"=>"1932-6203; 1932-6203"}, "id"=>"17810b08-83a7-3ec4-8c82-98541745010b", "abstract"=>"BACKGROUND: Recent progress in rat pluripotent stem cell technology has been remarkable. Particularly salient is the demonstration that embryonic stem cells (ESCs) in the rat (rESCs) can contribute to germline transmission, permitting generation of gene-modified rats as is now done using mouse ESCs (mESCs) or mouse induced pluripotent stem cells (iPSCs; miPSCs). However, determinations of whether rat iPSCs (riPSCs) can contribute to germ cells are not published. Here we report the germline competency of riPSCs.\\n\\nMETHODOLOGY/PRINCIPAL FINDINGS: We generated riPSCs by transducing three mouse reprogramming factors (Oct3/4, Klf4, and Sox2) into rat somatic cells, followed by culture in the presence of exogenous rat leukemia inhibitory factor (rLIF) and small molecules that specifically inhibit GSK3, MEK, and FGF receptor tyrosine kinases. We found that, like rESCs, our riPSCs can contribute to germline transmission. Furthermore we found, by immunostaining of testis from mouse-rat interspecific chimeras with antibody against mouse vasa homolog, that riPSCs can contribute to embryonic development with chimera formation in mice (rat-mouse interspecific chimeras) and to interspecific germlines.\\n\\nCONCLUSIONS/SIGNIFICANCE: Our data clearly demonstrate that using only three reprogramming factors (Oct3/4, Klf4, and Sox2) rat somatic cells can be reprogrammed into a ground state. Our generated riPSCs exhibited germline transmission in either rat-rat intraspecific or mouse-rat interspecific chimeras.", "link"=>"http://www.mendeley.com/research/generation-germlinecompetent-rat-induced-pluripotent-stem-cells", "reader_count"=>46, "reader_count_by_academic_status"=>{"Unspecified"=>1, "Professor > Associate Professor"=>6, "Researcher"=>10, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>2, "Other"=>1, "Student > Master"=>3, "Student > Bachelor"=>1, "Lecturer"=>3, "Professor"=>3}, "reader_count_by_user_role"=>{"Unspecified"=>1, "Professor > Associate Professor"=>6, "Researcher"=>10, "Student > Doctoral Student"=>5, "Student > Ph. D. Student"=>11, "Student > Postgraduate"=>2, "Other"=>1, "Student > Master"=>3, "Student > Bachelor"=>1, "Lecturer"=>3, "Professor"=>3}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>9, "Materials Science"=>2, "Medicine and Dentistry"=>8, "Agricultural and Biological Sciences"=>25}, "reader_count_by_subdiscipline"=>{"Materials Science"=>{"Materials Science"=>2}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>8}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>25}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>9}, "Unspecified"=>{"Unspecified"=>2}}, "reader_count_by_country"=>{"United States"=>1, "Portugal"=>1}, "group_count"=>0}

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Figshare

• {"files"=>["https://ndownloader.figshare.com/files/380349", "https://ndownloader.figshare.com/files/380379", "https://ndownloader.figshare.com/files/380436", "https://ndownloader.figshare.com/files/380473"], "description"=>"<div><h3>Background</h3><p>Recent progress in rat pluripotent stem cell technology has been remarkable. Particularly salient is the demonstration that embryonic stem cells (ESCs) in the rat (rESCs) can contribute to germline transmission, permitting generation of gene-modified rats as is now done using mouse ESCs (mESCs) or mouse induced pluripotent stem cells (iPSCs; miPSCs). However, determinations of whether rat iPSCs (riPSCs) can contribute to germ cells are not published. Here we report the germline competency of riPSCs.</p> <h3>Methodology/Principal Findings</h3><p>We generated riPSCs by transducing three mouse reprogramming factors (Oct3/4, Klf4, and Sox2) into rat somatic cells, followed by culture in the presence of exogenous rat leukemia inhibitory factor (rLIF) and small molecules that specifically inhibit GSK3, MEK, and FGF receptor tyrosine kinases. We found that, like rESCs, our riPSCs can contribute to germline transmission. Furthermore we found, by immunostaining of testis from mouse-rat interspecific chimeras with antibody against mouse vasa homolog, that riPSCs can contribute to embryonic development with chimera formation in mice (rat-mouse interspecific chimeras) and to interspecific germlines.</p> <h3>Conclusions/Significance</h3><p>Our data clearly demonstrate that using only three reprogramming factors (Oct3/4, Klf4, and Sox2) rat somatic cells can be reprogrammed into a ground state. Our generated riPSCs exhibited germline transmission in either rat-rat intraspecific or mouse-rat interspecific chimeras.</p> </div>", "links"=>[], "tags"=>["germline-competent", "induced", "pluripotent", "cells"], "article_id"=>135150, "categories"=>["Developmental Biology"], "users"=>["Sanae Hamanaka", "Tomoyuki Yamaguchi", "Toshihiro Kobayashi", "Megumi Kato-Itoh", "Satoshi Yamazaki", "Hideyuki Sato", "Ayumi Umino", "Yukiko Wakiyama", "Mami Arai", "Makoto Sanbo", "Masumi Hirabayashi", "Hiromitsu Nakauchi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022008.s001", "https://dx.doi.org/10.1371/journal.pone.0022008.s002", "https://dx.doi.org/10.1371/journal.pone.0022008.s003", "https://dx.doi.org/10.1371/journal.pone.0022008.s004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Generation_of_Germline_Competent_Rat_Induced_Pluripotent_Stem_Cells/135150", "title"=>"Generation of Germline-Competent Rat Induced Pluripotent Stem Cells", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-07-15 01:25:50"}
• {"files"=>["https://ndownloader.figshare.com/files/755794"], "description"=>"<p>(A) Schema of inducible lentivirus vector. XbaI enzyme restriction site exists between Ubiquitin-C (<i>Ubc</i>) promoter and reverse tet transactivator (rtTA) element. (B) Schematic time schedule of riPSCs generation. (C)–(D) Morphology of generated riPSCs (passage P0); bright field and fluorescent field (C). ALP staining after clonal passage (D).</p>", "links"=>[], "tags"=>["ips", "cells", "embryonic"], "article_id"=>426156, "categories"=>["Developmental Biology"], "users"=>["Sanae Hamanaka", "Tomoyuki Yamaguchi", "Toshihiro Kobayashi", "Megumi Kato-Itoh", "Satoshi Yamazaki", "Hideyuki Sato", "Ayumi Umino", "Yukiko Wakiyama", "Mami Arai", "Makoto Sanbo", "Masumi Hirabayashi", "Hiromitsu Nakauchi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022008.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Generation_of_rat_iPS_cells_from_rat_embryonic_fibroblasts_/426156", "title"=>"Generation of rat iPS cells from rat embryonic fibroblasts.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-15 01:42:36"}
• {"files"=>["https://ndownloader.figshare.com/files/755966"], "description"=>"<p>(A) Immunofluorescence staining for Oct3/4 and Nanog in rat iPS cells (riPSCs). (B) RT-PCR analysis of transgene <i>T2A-Sox2</i> and endogenous ES marker genes with or without doxycycline culture. RiPSC clones (riPSCs WI T3-11 and T1-3) express ESC markers. Rat and mouse embryonic fibroblast cells (REFs or MEFs) were used as negative controls. β-actin was used as a loading control. (C) Bisulfite genomic sequencing of the distal enhancer region of <i>Oct3/4</i>. Open and closed circles indicate unmethylated and methylated CpGs, respectively. (D) Hematoxylin/eosin staining of teratoma derived from rat iPS cells. Teratoma is composed of various types of tissues: Gut-like epithelium (endoderm), muscle and cartilage (mesoderm), epidermis, and neural tissue (ectoderm).</p>", "links"=>[], "tags"=>["ips"], "article_id"=>426332, "categories"=>["Developmental Biology"], "users"=>["Sanae Hamanaka", "Tomoyuki Yamaguchi", "Toshihiro Kobayashi", "Megumi Kato-Itoh", "Satoshi Yamazaki", "Hideyuki Sato", "Ayumi Umino", "Yukiko Wakiyama", "Mami Arai", "Makoto Sanbo", "Masumi Hirabayashi", "Hiromitsu Nakauchi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022008.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Characterization_of_rat_iPS_cells_/426332", "title"=>"Characterization of rat iPS cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-15 01:45:32"}
• {"files"=>["https://ndownloader.figshare.com/files/756175"], "description"=>"<p>(A) Cytogenetic analysis, riPSCs; G-band staining. Representative data (WI riPSC clone T3-11, passage 7) indicate a chromosomal number of 42, including an XY gender chromosome. (B) Efficiency of chimera rat generation in culture, with or without doxycycline. (C)–(D) Generation of chimeric rat. (C) Analysis of E15.5 material derived from WI riPSCs injected into WI blastocysts. EGFP indicates riPSC-derived cells. (D) Chimeric rats derived from DA riPSCs injected into WI blastocysts. Brown coat color indicates derivation from DA riPSCs. One-week-old pups; left pup (white) not chimeric (left panel). Three-month-old chimeric rat; eye color also is brown (right panel).</p>", "links"=>[], "tags"=>["chimeric", "derived", "ips"], "article_id"=>426531, "categories"=>["Developmental Biology"], "users"=>["Sanae Hamanaka", "Tomoyuki Yamaguchi", "Toshihiro Kobayashi", "Megumi Kato-Itoh", "Satoshi Yamazaki", "Hideyuki Sato", "Ayumi Umino", "Yukiko Wakiyama", "Mami Arai", "Makoto Sanbo", "Masumi Hirabayashi", "Hiromitsu Nakauchi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022008.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Generation_of_chimeric_rat_derived_from_rat_iPS_cells_/426531", "title"=>"Generation of chimeric rat derived from rat iPS cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-15 01:48:51"}
• {"files"=>["https://ndownloader.figshare.com/files/756359"], "description"=>"<p>(A) EGFP of offspring (E15.5; neonate) of chimeric rats. (B) FACS analysis of rat embryonic fibroblasts (REFs) established from F1 rats. (C) Genomic PCR, F1 rats. Some F1 rats harbored <i>EGFP</i>. Genomic DNA was obtained from F1 REFs or from tail. (D) Provirus copy numbers determined by Southern blot analysis. Lane1: WI riPSCs, clone T1-3; 2: WI riPSCs, clone T3-11; 3: DA riPSCs, clone T4-27; 4: DA riPSCs, clone T4-30; 5–7: EGFP-expressing F1 rats. (E) REFs established from F1 rats using Dox were reprogrammable: Re-generated riPSCs expressed EGFP and ALP.</p>", "links"=>[], "tags"=>["developmental biology"], "article_id"=>426723, "categories"=>["Developmental Biology"], "users"=>["Sanae Hamanaka", "Tomoyuki Yamaguchi", "Toshihiro Kobayashi", "Megumi Kato-Itoh", "Satoshi Yamazaki", "Hideyuki Sato", "Ayumi Umino", "Yukiko Wakiyama", "Mami Arai", "Makoto Sanbo", "Masumi Hirabayashi", "Hiromitsu Nakauchi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022008.g004"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Germline_transmission_of_riPSCs_/426723", "title"=>"Germline transmission of riPSCs.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-15 01:52:03"}
• {"files"=>["https://ndownloader.figshare.com/files/756566"], "description"=>"<p>(A) RiPSCs injected into mouse blastocysts; analysis, E13.5. EGFP of chimeric mouse (E13.5) generated using rat iPSCs. (B) EGFP (%) of chimerism was shown by FACS analysis of chimeric mouse embryonic fibroblasts (MEFs/REFs) established from F1 rats. (C) Immunostaining of chimeric rats. HMV/DTT (Alexa546) or EGFP fluorescence was observed in testis. EGFP was derived from riPSCs.</p>", "links"=>[], "tags"=>["interspecific", "chimeras"], "article_id"=>426929, "categories"=>["Developmental Biology"], "users"=>["Sanae Hamanaka", "Tomoyuki Yamaguchi", "Toshihiro Kobayashi", "Megumi Kato-Itoh", "Satoshi Yamazaki", "Hideyuki Sato", "Ayumi Umino", "Yukiko Wakiyama", "Mami Arai", "Makoto Sanbo", "Masumi Hirabayashi", "Hiromitsu Nakauchi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022008.g005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Generation_of_interspecific_chimeras_between_mouse_and_rat_/426929", "title"=>"Generation of interspecific chimeras between mouse and rat.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-07-15 01:55:29"}
• {"files"=>["https://ndownloader.figshare.com/files/756728"], "description"=>"<p>N/A: not applicable.</p><p>*1: One chimera died before weaning.</p>", "links"=>[], "tags"=>["ips"], "article_id"=>427093, "categories"=>["Developmental Biology"], "users"=>["Sanae Hamanaka", "Tomoyuki Yamaguchi", "Toshihiro Kobayashi", "Megumi Kato-Itoh", "Satoshi Yamazaki", "Hideyuki Sato", "Ayumi Umino", "Yukiko Wakiyama", "Mami Arai", "Makoto Sanbo", "Masumi Hirabayashi", "Hiromitsu Nakauchi"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0022008.t001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Chimera_rat_generation_from_rat_iPS_cells_/427093", "title"=>"Chimera rat generation from rat iPS cells.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-07-15 01:58:13"}

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F1000Prime