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 100ng/ml) or Noggin (250, 500 or 750ng/ml) or TGF-β1 (0, 0.1, 1 or 10ng/ml) or SB431542 (0, 10, 25 or 50μm), 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 1.0ng/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 50μm but not at other concentrations. The percentage increase in colony area was lower (p<0.05) with 10μm SB-431542 than that in the controls, whereas at higher concentrations of 25 or 50μm, 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.