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Supplementation of Vascular Endothelial Growth Factor during in vitro Maturation of Porcine Cumulus Oocyte Complexes and Subsequent Developmental Competence after Parthenogenesis and in vitro Fertilization

Authors: D. Biswas, Sang H. Hyun


In mammalian reproductive tract, the oviduct secretes huge number of growth factors and cytokines that create an optimal micro-environment for the initial stages of preimplantation embryos. Secretion of these growth factors is stage-specific. Among them, VEGF is a potent mitogen for vascular endothelium and stimulates vascular permeability. Apart from angiogenesis, VEGF in the oviduct may be involved in regulating the oocyte maturation and subsequent developmental process during embryo production in vitro. In experiment 1, to evaluate the effect of VEGF during IVM of porcine COC and subsequent developmental ability after PA and SCNT. The results from these experiments indicated that maturation rates among the different VEGF concentrations were not significant different. In experiment 2, total intracellular GSH concentrations of oocytes matured with VEGF (5-50 ng/ml) were increased significantly compared to a control and VEGF group (500 ng/ml). In experiment 3, the blastocyst formation rates and total cell number per blastocyst after parthenogenesis of oocytes matured with VEGF (5-50 ng/ml) were increased significantly compared to a control and VEGF group (500 ng/ml). Similarly, in experiment 4, the blastocyst formation rate and total cell number per blastocyst after SCNT and IVF of oocytes matured with VEGF (5 ng/ml) were significantly higher than that of oocytes matured without VEGF group. In experiment 5, at 10 hour after the onset of IVF, pronuclear formation rate was evaluated. Monospermy was significantly higher in VEGF-matured oocytes than in the control, and polyspermy and sperm penetration per oocyte were significantly higher in the control group than in the VEGFmatured oocytes. Supplementation with VEGF during IVM significantly improved male pronuclear formation as compared with the control. In experiment 6, type III cortical granule distribution in oocytes was more common in VEGF-matured oocytes than in the control. In conclusion, the present study suggested that supplementation of VEGF during IVM may enhance the developmental potential of porcine in vitro embryos through increase of the intracellular GSH level, higher MPN formation and increased fertilization rate as a consequence of an improved cytoplasmic maturation.

Keywords: angiogenesis, VEGF, GSH, monospermy

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[1] K.S. Richter, The importance of growth factors for preimplantation embryo development and in vitro culture, Curr. Opin. Obstet. Gynecol., vol. 20, pp. 292-304, 2008.
[2] A.J. Watson, Oocyte cytoplasmic maturation: A key mediator of oocyte and embryo developmental competence, J. Anim. Sci.,vol. 85, pp. E1- E3, 2007.
[3] H. Luo, K. Kimura, M. Aoki and M. Hirako, Effect of Vascular Endothelial Growth Factor on Maturation, Fertilization and Developmental Competence of Bovine Oocytes, J. Vet. Med. Sci., vol, 64, pp. 803-806, 2002.
[4] J. Tran, Z. Master, J.L. Yu, J. Rak, D.J. Dumont and R.S. Kerbel, A role for surviving in chemoresistance of endothelial cells mediated by VEGF, PNAS, USA, vol, 99, pp. 4349-4354, 2002.
[5] H. Funahashi, T.C. Cantley, T.T. Stumpf, S.L. Terlouw and B.N. Day, Use of low-salt culture medium for in vitro maturation of porcine oocytes is associated with elevated oocyte glutathione levels and enhanced male pronuclear formation after in vitro fertilization, Biol. Reprod., vol. 51, pp. 633-639, 1994.
[6] L. Zofia, The role of glutathione in mammalian gametes, Reproductive Biol., vol. 5, no. 1, pp. 5-17, 2005.
[7] O.J. Koo, G. Jang, D.K. Kwon, J.T. Kang, O.S. Kwon, H.J. Park, S.K. Kang and B.C. Lee, Electrical activation induces reactive oxygen species in porcine embryos, Theriogenology, vol. 70, no. 7, pp. 1111-1118, 2008.
[8] P.J.Booth, S.J. Tan, R. Reipurth, P. Holm and H. Callesen, Simplification of bovine somatic cell nuclear transfer by application of a zona-free manipulation technique, Clon. Stem Cells, vol. 3, pp. 139-150, 2001.
[9] W.J. Jolliff and R.S. Prather, Parthenogenic development of in vitro matured, in vivo-cultured porcine oocytes beyond blastocyst, Biol. Reprod., vol. 56, pp. 544-548, 1997.
[10] L.R Abeydeera, In vitro fertilization and embryo development in pigs. Reprod., vol. 58, pp. 159-173, 2001.
[11] A.C. Boquest, L.R. Abeydeera, W.H. Wang and B.N. Day, Effect of adding reduced glutathione during insemination on the development of porcine embryos in vitro, Theriogenology, vol. 51, pp. 1311-1319, 1999.
[12] K.A. Zuelke, S.C. Jeffay, R.M. Zucker, S.D. Perreault, Glutathione (GSH) concentrations vary with the cell cycle in maturing hamster oocytes, zygotes, and pre-implantation stage embryos, Mol. Reprod.Deve., vol. 64, pp. 106-112, 2003.
[13] H.I. Calvin, K. Grosshans and E.J. Blake, Estimation and manipulation of glutathione levels in prepuberal mouse ovaries and ova: relevance to sperm nucleus transformation in the fertilized egg, Gamete Res., vol. 14, pp. 265-275, 1986.
[14] SD Perreault, RR Barbee, VL Slott, Importance of glutathione in the acquisition and maintenance of sperm nuclear decondensing activity in maturing hamster oocytes, Deve. Biol., vol. 125, pp. 181-186, 1988.
[15] M. Yoshida, Role of glutathione in the maturation and fertilization of pig oocytes in vitro, Mol. Reprod. Deve., vol. 35, pp. 76-81, 1993.
[16] M.H. Nasr-Esfahani and M.H. Johnson, Quantitative analysis of cellular glutathione in early preimplantation mouse embryos developing in vivo and in vitro, Hum.Reprod., vol. 7, pp. 1281-1290, 1992
[17] KS Viana, MC Caldas-Bussiere, SG Matta, MR Faes, CS Paes de Carvalho, CR Quirino, Effect of sodium nitroprusside, a nitric oxide donor, on the in vitro maturation of bovine oocytes, Anim. Reprod. Sci., vol. 102, pp. 217-227, 2006.