title: Production of feline leukemia inhibitory factor with biological activity in Escherichia coli.
abstract: Leukemia inhibitory factor (LIF) is a cytokine which is essential for oocyte and embryo development, embryonic stem cell, and induced pluripotent stem cell maintenance. Leukemia inhibitory factor improves the maturation of oocytes in the human and the mouse. However, feline LIF (fLIF) cloning and effects on oocytes during IVM have not been reported. Thus, we cloned complete cDNA of fLIF and examined its biological activity and effects on oocytes during IVM in the domestic cat. The aminoacid sequence of fLIF revealed a homology of 81% or 92% with that of mouse or human. The fLIF produced by pCold TF DNA in Escherichia coli was readily soluble and after purification showed bioactivity in maintaining the undifferentiated state of mouse embryonic stem cells and enhancing the proliferation of human erythrocyte leukemia cells. Furthermore, 10- and 100-ng/mL fLIF induced cumulus expansion with or without FSH and EGF (P < 0.05). The rate of metaphase II oocytes was also improved with 100-ng/mL fLIF (P < 0.05). We therefore confirmed the successful production for the first time of biologically active fLIF and revealed its effects on oocytes during IVM in the domestic cat. Feline LIF will further improve reproduction and stem cell research in the feline family.
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Kanegi
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Hatoya
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Tsujimoto
authors.affiliation: Department of Integrated Functional Biosciences, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Takenaka
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Nishimura
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan.
authors.forename: V
authors.initials: V
authors.lastname: Wijewardana
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Sugiura
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Takahashi
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Kawate
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Tamada
authors.affiliation: Department of Advanced Pathobiology, Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Izumisano, Osaka, Japan. Electronic address: inaba@vet.osakafu-u.ac.jp.
authors.forename: T
authors.initials: T
authors.lastname: Inaba
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002415:Cats / Q000502:physiology*; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004622:Embryo, Mammalian / Q000166:cytology; D004926:Escherichia coli / Q000378:metabolism*; D005347:Fibroblasts / Q000502:physiology; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D053605:Leukemia Inhibitory Factor / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids
chemical_list: D018076:DNA, Complementary; D053605:Leukemia Inhibitory Factor
keywords: Cumulus expansion; Feline; IVM; Leukemia inhibitory factor; Nuclear maturation; Recombinant protein