title: Derivation of embryonic stem cell lines from parthenogenetically developing rat blastocysts.
abstract: This study was undertaken to establish rat embryonic stem (ES) cells from parthenogenetically developing blastocysts. Ten blastocysts were prepared by treatment of ovulated rat oocytes with ionomycin and cycloheximide, and three alkaline phosphatase-positive ES cell lines were established using the N2B27 medium supplemented with mitogen activated protein kinase kinase inhibitor PD0325901, glycogen synthase kinase 3 inhibitor CHIR99021, rat leukemia inhibitory factor, and forskolin. Expression of stem cell marker genes (Oct-4, rNanog, Fgf-4, and Rex-1) was confirmed in all three ES cell lines by reverse transcriptase-polymerase chain reaction (RT-PCR). Combined bisulfite restriction analysis showed that the differentially methylated region locus of five imprinted genes (H19, Meg3IG, Igf2r, Peg5, and Peg10) in these ES cells remained to be demethylated or was hypomethylated, which was similar to that in control ES cells established from normal blastocysts. Characteristics of the parthenogenetic blastocyst-derived ES cells were successfully transmitted to the next generation through a chimeric rat for one of the three ES cell lines. This is the first report on germline-competent (genuine) ES cells derived from parthenogenetically developing rat blastocysts.
authors.affiliation: 1 Section of Mammalian Transgenesis, Center for Genetic Analysis of Behavior, National Institute for Physiological Sciences , Aichi, Japan .
authors.forename: Masumi
authors.initials: M
authors.lastname: Hirabayashi
authors.forename: Teppei
authors.initials: T
authors.lastname: Goto
authors.forename: Chihiro
authors.initials: C
authors.lastname: Tamura
authors.forename: Makoto
authors.initials: M
authors.lastname: Sanbo
authors.forename: Hiromasa
authors.initials: H
authors.lastname: Hara
authors.forename: Megumi
authors.initials: M
authors.lastname: Kato-Itoh
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Sato
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Kobayashi
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Nakauchi
authors.forename: Shinichi
authors.initials: S
authors.lastname: Hochi
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D001755:Blastocyst / Q000378:metabolism*; D061207:Calcium Ionophores / Q000494:pharmacology; D002454:Cell Differentiation; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D019175:DNA Methylation; D004159:Diphenylamine / Q000031:analogs & derivatives / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000378:metabolism*; D005260:Female; D051519:Fibroblast Growth Factor 4 / Q000096:biosynthesis; D005819:Genetic Markers; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors; D015759:Ionomycin / Q000494:pharmacology; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology; D020929:Mitogen-Activated Protein Kinase Kinases / Q000037:antagonists & inhibitors; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000096:biosynthesis; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism*; D010312:Parthenogenesis; D011725:Pyridines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D051381:Rats; D014157:Transcription Factors / Q000096:biosynthesis
chemical_list: D000276:Adjuvants, Immunologic; D001549:Benzamides; D061207:Calcium Ionophores; C473711:Chir 99021; C496447:Fgf4 protein, rat; D051519:Fibroblast Growth Factor 4; D005819:Genetic Markers; D053605:Leukemia Inhibitory Factor; D000071317:Nanog Homeobox Protein; C547030:Nanog protein, rat; D050814:Octamer Transcription Factor-3; D011725:Pyridines; D011743:Pyrimidines; D014157:Transcription Factors; D005576:Colforsin; D015759:Ionomycin; C506614:mirdametinib; D003513:Cycloheximide; D004159:Diphenylamine; D038362:Glycogen Synthase Kinase 3; D020929:Mitogen-Activated Protein Kinase Kinases