title: LIF and FGF cooperatively support stemness of rabbit embryonic stem cells derived from parthenogenetically activated embryos.
abstract: We investigated the individual and combined effects of leukemia inhibitory factor (LIF) and basic fibroblast growth factor 2 (bFGF2) on the derivation and maintenance of rabbit embryonic stem cell lines isolated from parthenogenetic activated embryos (p-rES). First, we demonstrated that p-rES cell lines can be prevented from differentiation via LIF (STAT3) and bFGF2 (MEK-ERK1/2 and PI3K-AKT) signaling on MEF feeders. High levels of ERK1/2 and AKT activities were crucial for maintaining p-rES cells in an undifferentiated state. Although the p-rES cells under the influence of LIF (500, 1000, and 2000 U/mL) or bFGF2 (5, 10, and 20 ng/mL) alone showed enhanced expression in the pluripotency markers, the highest levels of marker expressions coincided with the simultaneous presence of LIF (1000 U/mL) and bFGF2 (10 ng/mL). The phosphorylation status of LIF and bFGF2 downstream signaling molecules including STAT3, ERK, and AKT was also intensively involved in the maintenance of p-rES cell proliferation and self-renewal. Induced dephosphorylation of STAT3, ERK1/2, and AKT by specific inhibitors caused remarkable losses of self-renewal capacity of p-rES cells. We conclude that bFGF2 and LIF by itself are self-sufficient in maintaining the state of undifferentiation and self-renewal of rabbit p-ES cells, yet are most effective when acting concomitantly.
authors.affiliation: Department of Animal Science, National Chung Hsing University, Taichung, Taiwan, Republic of China.
authors.forename: Ya-Chen
authors.initials: YC
authors.lastname: Hsieh
authors.forename: Payungsuk
authors.initials: P
authors.lastname: Intawicha
authors.forename: Kun-Hsiung
authors.initials: KH
authors.lastname: Lee
authors.forename: Yung-Tsung
authors.initials: YT
authors.lastname: Chiu
authors.forename: Neng-Wen
authors.initials: NW
authors.lastname: Lo
authors.forename: Jyh-Cherng
authors.initials: JC
authors.lastname: Ju
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D048052:Mitogen-Activated Protein Kinase 3 / Q000502:physiology; D023421:Models, Animal; D010312:Parthenogenesis / Q000502:physiology*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D051057:Proto-Oncogene Proteins c-akt / Q000502:physiology; D011817:Rabbits; D050796:STAT3 Transcription Factor / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology
chemical_list: D053605:Leukemia Inhibitory Factor; D050796:STAT3 Transcription Factor; D016222:Fibroblast Growth Factor 2; D051057:Proto-Oncogene Proteins c-akt; D048052:Mitogen-Activated Protein Kinase 3; D000469:Alkaline Phosphatase