Parthenogenic development of in vitro-matured, in vivo-cultured porcine oocytes beyond blastocyst.
Some of the factors influencing the success of a nuclear transfer procedure are the quality of the recipient oocyte and the efficiency of the method of artificial activation. In this study we evaluated the ability of an electrical pulse to stimulate in vitro-matured porcine oocytes to develop. Maturation in Waymouth's medium resulted in significantly greater development than maturation in TCM-199 (p < 0.05), while there was no significant difference between degrees of development in Waymouth's medium and Whitten's medium. Oocytes matured in Waymouth's medium and electrically stimulated at 36 h (young oocytes) developed to the same degree as oocytes stimulated at 48 h (aged oocytes). Oocytes matured in Waymouth's medium and treated with cytochalasin B showed significantly greater development (p < 0.10) in response to electrical activation than controls. Staurosporine activation of oocytes resulted in significantly (p < 0.05) fewer morulae and blastocysts when compared to electrical stimulation. Development of parthenogenic embryos to the elongated filamentous stage (10% development beyond blastocyst) was obtained by maturing oocytes in Waymouth's medium and electrically stimulating them to develop. By obtaining development of porcine parthenotes beyond the blastocyst stage, we have identified an efficient method of oocyte maturation and oocyte activation for use in a system for nuclear transfer.
Jolliff
W J
WJ
Department of Animal Sciences, University of Missouri-Columbia, 65211, USA.
Prather
R S
RS
D000818:Animals; D001755:Blastocyst / Q000502:physiology*; D002478:Cells, Cultured; D003470:Culture Media; D004558:Electric Stimulation; D005314:Embryonic and Fetal Development; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D009028:Morula / Q000502:physiology; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000187:drug effects / Q000502:physiology*; D010312:Parthenogenesis; D047428:Protein Kinase Inhibitors; D019311:Staurosporine / Q000494:pharmacology; D013552:Swine; D013997:Time Factors
D003470:Culture Media; D004791:Enzyme Inhibitors; D047428:Protein Kinase Inhibitors; D019311:Staurosporine