RRC ID 51645
Author Kanegi R, Hatoya S, Tsujimoto Y, Takenaka S, Nishimura T, Wijewardana V, Sugiura K, Takahashi M, Kawate N, Tamada H, Inaba T.
Title Production of feline leukemia inhibitory factor with biological activity in Escherichia coli.
Journal Theriogenology
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.
Volume 86(2)
Pages 604-11
Published 2016-7-15
DOI 10.1016/j.theriogenology.2016.02.013
PII S0093-691X(16)00085-6
PMID 27020881
MeSH Amino Acid Sequence Animals Base Sequence Cats / physiology* Cloning, Molecular DNA, Complementary Embryo, Mammalian / cytology Escherichia coli / metabolism* Fibroblasts / physiology Gene Expression Regulation, Bacterial / physiology Leukemia Inhibitory Factor / genetics Leukemia Inhibitory Factor / metabolism* Plasmids
IF 2.094
Times Cited 4
Human and Animal Cells TT2(AES0014)