title: Derivation, characterization, and stable transfection of induced pluripotent stem cells from Fischer344 rats.
abstract: 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.
authors.affiliation: Institute of Cytology, Russian Academy of Sciences, St. Petersburg, Russia.
authors.forename: Mikhail
authors.initials: M
authors.lastname: Liskovykh
authors.forename: Ilya
authors.initials: I
authors.lastname: Chuykin
authors.forename: Ashish
authors.initials: A
authors.lastname: Ranjan
authors.forename: Dina
authors.initials: D
authors.lastname: Safina
authors.forename: Elena
authors.initials: E
authors.lastname: Popova
authors.forename: Elena
authors.initials: E
authors.lastname: Tolkunova
authors.forename: Valentina
authors.initials: V
authors.lastname: Mosienko
authors.forename: Julia M
authors.initials: JM
authors.lastname: Minina
authors.forename: Natalia S
authors.initials: NS
authors.lastname: Zhdanova
authors.forename: John J
authors.initials: JJ
authors.lastname: Mullins
authors.forename: Michael
authors.initials: M
authors.lastname: Bader
authors.forename: Natalia
authors.initials: N
authors.lastname: Alenina
authors.forename: Alexey
authors.initials: A
authors.lastname: Tomilin
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D018274:Electroporation; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D018390:Gene Targeting; D005822:Genetic Vectors; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007124:Immunoenzyme Techniques; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000502:physiology*; D007621:Karyotyping; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D011916:Rats, Inbred F344; D013724:Teratoma / Q000378:metabolism / Q000473:pathology*; D014162:Transfection
chemical_list: D004791:Enzyme Inhibitors; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D048049:Extracellular Signal-Regulated MAP Kinases; D038362:Glycogen Synthase Kinase 3; D020929:Mitogen-Activated Protein Kinase Kinases; D000469:Alkaline Phosphatase
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