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Successful Culture In Vitro of Swine Embryos to the Blastocyst Stage^{1 ,2,}

Washington State University³, Pullman 99163

Abstract

One hundred and twenty-three porcine embryos between the two-cell and morula stage of development were collected from 24 gilts. Embryos were placed randomly into culture among three media and two gaseous atmospheres and development was recorded at 24-hr intervals for the duration of culture. The number of cells present at the cessation of culture was determined following aceto-orcein staining.

Development of embryos in vitro was measured by a cleavage index based on the number of cells present at the cessation of culture. Significant differences were found among the culture media and gaseous atmospheres with a significant (P<.02) media x gaseous atmosphere interaction. Embryos in Whitten's medium (WM) with 1 mg/ml bovine serum albumin (BSA), WM with 15 mg/ml (BSA) or Modified Ham's F-10 (MHF-10) all cultured under a 5% C0₂ in air atmosphere had cleavage indices of .69, 1.00 and .53, respectively. Embryos in WM with 1 mg/ml BSA, WM with 15 mg/ml BSA, or MHF-10 all cultured under a reduced gaseous atmosphere of 90% N₂, 5% CO₂ and 5% O₂, had cleavage indices of 1.43, 1.93 and .71, respectively. The stages of development at the time of initiation of culture influenced (P<.001) the degree to which the embryos ultimately developed when development was measured by a cleavage index. Five-to seven-cell (2/23), eight to 12-cell (6/28), andmorulae (19/20) developed into blastocysts with one to two-cell and three to four-cell embryos failing to develop past the morula stage.

¹ Scientific Paper No. 4670. College of Agriculture Research Center, Washington State University, Pull man. Project 0313.

² The author wishes to thank the Washington State Swine Breeders Association for providing the gilts and feed used in this study, Dr. C. T. Gaskins for statistical assistance, and Drs. J. A. Froseth and E. L. Martin for contributions in this experiment.

³ Department of Animal Sciences.