title: Effect of TGF-β1 superfamily members on survival of buffalo (Bubalus bubalis) embryonic stem-like cells.
abstract: This study examined the effects of supplementation of ES-like cell culture medium with bone morphogenetic protein (BMP)-4 (0, 10, 20 or 100 ng/ml) or Noggin (250, 500 or 750 ng/ml) or TGF-β1 (0, 0.1, 1 or 10 ng/ml) or SB431542 (0, 10, 25 or 50 μm), an inhibitor of TGF-β1 signalling, on survival, colony area and expression level of pluripotency genes in buffalo ES-like cells at passage 40-80, under different culture conditions. BMP-4 supplementation significantly reduced (p < 0.05) colony survival rate, percentage increase in colony area and relative mRNA abundance of OCT4, whereas that of NANOG and SOX-2 was increased significantly (p < 0.05). Noggin supplementation did not affect the colony survival rate and percentage increase in colony area in the presence of FGF-2 and LIF. In the presence of FGF-2 alone, it significantly reduced (p < 0.05) the relative mRNA abundance of OCT4 and SOX-2 and increased (p < 0.05) that of NANOG. Supplementation with TGF-β1 at 1.0 ng/ml but not at other concentrations increased colony survival rate but had no effect on percentage increase in colony area at any concentration. Supplementation with SB-431542 decreased (p < 0.05) colony survival rate at 50 μm but not at other concentrations. The percentage increase in colony area was lower (p < 0.05) with 10 μm SB-431542 than that in the controls, whereas at higher concentrations of 25 or 50 μm, SB-431542 decreased (p < 0.05) the colony size instead of increasing it. In conclusion, these results suggest that BMP-4 induces differentiation in buffalo ES-like cells, whereas TGF-β/activin/nodal pathway may not be playing a crucial role in maintaining pluripotency in these cells.
authors: Embryo Biotechnology Lab, Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana, India.
authors: R
authors: R
authors: Sharma
authors: N M
authors: NM
authors: Kamble
authors: A
authors: A
authors: George
authors: M S
authors: MS
authors: Chauhan
authors: S
authors: S
authors: Singla
authors: R S
authors: RS
authors: Manik
authors: P
authors: P
authors: Palta
mesh_terms: D000818:Animals; D001549:Benzamides / Q000008:administration & dosage; D019485:Bone Morphogenetic Proteins / Q000008:administration & dosage; D002020:Buffaloes / Q000196:embryology*; D002352:Carrier Proteins / Q000008:administration & dosage; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D004149:Dioxoles / Q000008:administration & dosage; D053595:Embryonic Stem Cells / Q000502:physiology*; D016222:Fibroblast Growth Factor 2 / Q000008:administration & dosage; D053605:Leukemia Inhibitory Factor / Q000008:administration & dosage; D053773:Transforming Growth Factor beta1 / Q000008:administration & dosage* / Q000037:antagonists & inhibitors; D000099176:Noggin Protein
chemical_list: D001549:Benzamides; D019485:Bone Morphogenetic Proteins; D002352:Carrier Proteins; D003470:Culture Media; D004149:Dioxoles; D016222:Fibroblast Growth Factor 2; D053605:Leukemia Inhibitory Factor; D053773:Transforming Growth Factor beta1; D000099176:Noggin Protein; C459179:4-(5-benzo(1,3)dioxol-5-yl-4-pyridin-2-yl-1H-imidazol-2-yl)benzamide