Derivation, Characterization, and Stable Transfection of Induced Pluripotent Stem Cells from Fischer344 Rats
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Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/362839", "https://ndownloader.figshare.com/files/362887", "https://ndownloader.figshare.com/files/362979", "https://ndownloader.figshare.com/files/363016", "https://ndownloader.figshare.com/files/363118", "https://ndownloader.figshare.com/files/363176", "https://ndownloader.figshare.com/files/363215", "https://ndownloader.figshare.com/files/363275"], "description"=>"<div><p>The rat represents an important animal model that, in many respects, is superior to the mouse for dissecting behavioral, cardiovascular and other physiological pathologies relevant to humans. Derivation of induced pluripotent stem cells from rats (riPS) opens the opportunity for gene targeting in specific rat strains, as well as for the development of new protocols for the treatment of different degenerative diseases. Here, we report an improved lentivirus-based hit-and-run riPS derivation protocol that makes use of small inhibitors of MEK and GSK3. We demonstrate that the excision of proviruses does not affect either the karyotype or the differentiation ability of these cells. We show that the established riPS cells are readily amenable to genetic manipulations such as stable electroporation. Finally, we propose a genetic tool for an improvement of riPS cell quality in culture. These data may prompt iPS cell-based gene targeting in rat as well as the development of iPS cell-based therapies using disease models established in this species.</p> </div>", "links"=>[], "tags"=>["transfection", "induced", "pluripotent", "cells", "fischer344", "rats"], "article_id"=>131707, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Mikhail Liskovykh", "Ilya Chuykin", "Ashish Ranjan", "Dina Safina", "Elena Popova", "Elena Tolkunova", "Valentina Mosienko", "Julia M. Minina", "Natalia S. Zhdanova", "John J. Mullins", "Michael Bader", "Natalia Alenina", "Alexey Tomilin"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0027345.s001", "https://dx.doi.org/10.1371/journal.pone.0027345.s002", "https://dx.doi.org/10.1371/journal.pone.0027345.s003", "https://dx.doi.org/10.1371/journal.pone.0027345.s004", "https://dx.doi.org/10.1371/journal.pone.0027345.s005", "https://dx.doi.org/10.1371/journal.pone.0027345.s006", "https://dx.doi.org/10.1371/journal.pone.0027345.s007", "https://dx.doi.org/10.1371/journal.pone.0027345.s008"], "stats"=>{"downloads"=>31, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/Derivation_Characterization_and_Stable_Transfection_of_Induced_Pluripotent_Stem_Cells_from_Fischer344_Rats/131707", "title"=>"Derivation, Characterization, and Stable Transfection of Induced Pluripotent Stem Cells from Fischer344 Rats", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2011-11-04 00:28:27"}
  • {"files"=>["https://ndownloader.figshare.com/files/716163"], "description"=>"<p>*Subclones of IIIB9 following Cre-mediated lentivirus excision.</p>", "links"=>[], "tags"=>["karyotype", "established", "rips"], "article_id"=>386513, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Mikhail Liskovykh", "Ilya Chuykin", "Ashish Ranjan", "Dina Safina", "Elena Popova", "Elena Tolkunova", "Valentina Mosienko", "Julia M. Minina", "Natalia S. Zhdanova", "John J. Mullins", "Michael Bader", "Natalia Alenina", "Alexey Tomilin"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027345.t002", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Summary_of_karyotype_analysis_of_established_riPS_cell_lines_/386513", "title"=>"Summary of karyotype analysis of established riPS cell lines.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-11-04 01:48:33"}
  • {"files"=>["https://ndownloader.figshare.com/files/716059"], "description"=>"<p>. riPS cells (here clone H5) harboring the <i>2A2Btk-TKiresPuro</i> transgenic cassette show an increasing percentage of spontaneously differentiated cells with flat morphology and reduced alkaline phosphatase (AP) expression with increasing number of passages in N2B27+2i+LIF medium (left column), whereas the differentiated cells have been mostly eliminated from the culture following a 3-day selection in the same culture medium containing 1 µg/ml puromycin (right column). PH – phase contrast.</p>", "links"=>[], "tags"=>["undifferentiated", "rips", "cells"], "article_id"=>386405, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Mikhail Liskovykh", "Ilya Chuykin", "Ashish Ranjan", "Dina Safina", "Elena Popova", "Elena Tolkunova", "Valentina Mosienko", "Julia M. Minina", "Natalia S. Zhdanova", "John J. Mullins", "Michael Bader", "Natalia Alenina", "Alexey Tomilin"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027345.g003", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Positive_selection_of_undifferentiated_riPS_cells_in_vitro_/386405", "title"=>"Positive selection of undifferentiated riPS cells <i>in vitro</i>", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-04 01:46:45"}
  • {"files"=>["https://ndownloader.figshare.com/files/715915"], "description"=>"<p>(<b>A</b>) Immunostaining of cultured riPS cells for Nanog (left) (63x objective, confocal imaging, clone IVC2), Oct4 (middle, clone IIIB9), and SSEA1 (right, clone G4) (20x objective, fluorescent microscope). (<b>B</b>) RT-PCR analysis of gene expression in undifferentiated riPS cells (clone H5) and in cells during the course of <i>in vitro</i> differentiation via the embryoid body (EB) protocol after 2 days (EB2), 6 days (EB6) and 12 days (dpp6, 6 days post plating) of differentiation; shown is the analysis of mesodermal (<i>AFP, Flk1</i>), endodermal (<i>Gata4</i> and <i>Sox17</i>), and ectodermal marker (<i>Nestin</i>) expression. (<b>C</b>) Differentiation of riPS cells into neurons <i>in vitro</i>, shown by immunostaining with Tuj1 antibodies at dpp6 of differentiation (clone H5). (<b>D</b>) H&E staining of sections of teratomas formed after a subcutaneous injection of riPS cells (clone G3) into NUDE rats; shown are stratified epithelium (left), cartilage (middle), and epithelium, likely of endodermal origin (right), belonging to ectodermal, mesodermal, and endodermal germ layers, respectively. (<b>E</b>) riPS cells (clone H5) injected into 8-cell rat embryos can contribute to the formation of the embryo proper and only to a very limited extend to the placenta, as shown by PCR analysis of day 18 rat embryo DNA. H5 – riPS cell clone used for the injection, 12 and 13 – positive and negative fetuses, respectively. T – tail, L – liver, H – head, K – kidney, P – placenta.</p>", "links"=>[], "tags"=>["generated", "rips"], "article_id"=>386262, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Mikhail Liskovykh", "Ilya Chuykin", "Ashish Ranjan", "Dina Safina", "Elena Popova", "Elena Tolkunova", "Valentina Mosienko", "Julia M. Minina", "Natalia S. Zhdanova", "John J. Mullins", "Michael Bader", "Natalia Alenina", "Alexey Tomilin"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027345.g002", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Pluripotency_of_generated_riPS_cells_/386262", "title"=>"Pluripotency of generated riPS cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-04 01:44:22"}
  • {"files"=>["https://ndownloader.figshare.com/files/716209"], "description"=>"<p>To derive rat iPS (riPS) cells 1.5×10<sup>5</sup> REFs were transduced with the LVTHM-based Oct4, Sox2, Klf4, cMyc, and EGFP lentiviruses. Four days later cells were split onto a 10 cm culture dish and cultured in one of the three reprogramming media. After 10–12 days, primary riPS cell colonies were counted to calculate the reprogramming efficiency. Survival rate was calculated as a relation between the number of picked and the number of surviving clones after 2 passages.</p><p>*p<0.05, Student's <i>t</i>-test condition 3 <i>vs</i> condition 2, [DMEM + SR +2i + LIF + A-83-01] N = 3; [N2B27+2i + LIF]: N = 4 independent experiments.</p>", "links"=>[], "tags"=>["rips", "derivation"], "article_id"=>386550, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Mikhail Liskovykh", "Ilya Chuykin", "Ashish Ranjan", "Dina Safina", "Elena Popova", "Elena Tolkunova", "Valentina Mosienko", "Julia M. Minina", "Natalia S. Zhdanova", "John J. Mullins", "Michael Bader", "Natalia Alenina", "Alexey Tomilin"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027345.t001", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Efficiencies_of_riPS_cell_derivation_in_different_culture_conditions_/386550", "title"=>"Efficiencies of riPS cell derivation in different culture conditions.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2011-11-04 01:49:10"}
  • {"files"=>["https://ndownloader.figshare.com/files/715762"], "description"=>"<p>(<b>A</b>) Rat embryonic fibroblasts (REFs) (left), primary riPS cell colony derived thereof after transduction with lentiviruses expressing <i>Oct4</i>, <i>Klf4</i>, <i>Sox2</i>, <i>cMyc</i>, and <i>EGFP</i> with some satellite colonies (middle), and riPS cell line after several passages in N2B27+LIF+2i medium (right). (<b>B</b>) Karyotyping results of the G4 riPS cell clone, in which 90% of metaphase plates showed the normal male chromosomal content (42,XY). (<b>C</b>) Cre-mediated excision of the proviruses in riPS cell clones H5, G3, and G4 from the genome of the parental IIIB9 riPS cell clone, shown by PCR of genomic DNA with primers specific for exogenous <i>Oct4</i>, <i>Klf4</i>, <i>Sox2</i>, <i>cMyc</i>, and <i>EGFP</i> sequences. PCR for the Y-chromosome specific gene Sry confirmed the DNA integrity and male origin of riPS clone IIIB9. (<b>D</b>) Cre-mediated excision of EGFP-lentivirus visualized in G3 riPS cell clone under fluorescent microscope.</p>", "links"=>[], "tags"=>["ips"], "article_id"=>386111, "categories"=>["Biochemistry", "Cell Biology", "Developmental Biology"], "users"=>["Mikhail Liskovykh", "Ilya Chuykin", "Ashish Ranjan", "Dina Safina", "Elena Popova", "Elena Tolkunova", "Valentina Mosienko", "Julia M. Minina", "Natalia S. Zhdanova", "John J. Mullins", "Michael Bader", "Natalia Alenina", "Alexey Tomilin"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0027345.g001", "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Generation_of_rat_iPS_riPS_cells_/386111", "title"=>"Generation of rat iPS (riPS) cells.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2011-11-04 01:41:51"}

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Relative Metric

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