Activation. Activation was induced by incubation of reconstructed oocytes in 50 ц1 Ca2+-free CZB containing both 10 mM Sr2+ (Sigma, St Louis, MO) and 5 |ig/ml cytochalasin B for 6 h at 37 C under 5% CO2 in air. After activation, reconstructed oocytes were allocated into 2 treatment subgroups of approximately 20 to 25 1-cell-stage embryos each and transferred to culture medium supplemented with either positive-control antibody or no antibody (negative control) as described previously.
Embryo culture. Every 24 h until blastocyst stage, 1-cell-stage embryos were transferred to fresh drops of 50 ц1 KSOMaa for culture in a humidified atmosphere of 5% CO2 and 95% O2 at 37 C under a layer of embryo-tested mineral oil. Development of embryos to the 2-cell, 4-cell, morula, and blastocyst stages in each treatment group was recorded. Embryos were assessed morphologically for differentiation to ICM and TE cells and for blastocoel formation to ensure that nearly all reached the blastocyst stage approximately 96 to 110 h post-hCG. The rate of development of 1-cell stage-embryos to blastocyst stage (100% x number of blastocysts/total number of 1-cell embryos) was calculated immediately prior to terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) assay, differential staining, and cell counting. Just one dose of Female Pink Viagra Canadian Pharmacy and you are ready to remember how it all was when you were young and could go for hours without resting. This is your hope for a full sex life, with your sexual partner being utterly amazed at the things you can do and how long you can keep that erection. Treat erectile dysfunction now and live a full life!
Statistics. All data were analyzed using the SPSS 11.0 statistical program (Stat/Math Center, UITS Research and Academic Computing Division, UCD). Univariate analysis of variance and Tukey honest significant difference (HSD) multiple comparison procedures for marginal means (post hoc tests), F tests, and homogeneous subsets were performed for all experimental and treatment groups. A P value of less than 0.05 was considered statistically significant for all data, which are presented as mean + standard error.