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• W2013301740 abstract "ObjectiveDetermine effects of single versus sequential culture media on murine embryo quality and developmental attributes, which are not accessible in human embryos.DesignMurine embryos cultured in commercially available single and sequential media were used to identify optimal preimplantation embryo quality and development attributes.Materials and MethodsMurine zygotes (n=40) were cultured in commercially available single or sequential media for 5 days. Images (200x, 400x) made using inverted phase contrast microscopes with differential contrast on developmental day 3 (d3) to facilitate quantification of the number of embryos developing to the 8-cell stage or greater in the two media. On d5, images were made and embryos fixed. Comparing and contrasting embryo images from the two types of media facilitated identification of an inner cell mass (ICM), nuclear quality assessment, and quantification of the total number of cells per embryo, ICM and trophoblast. The least square means (LSM) of total nuclei number, ICM nuclear number, and presumptive TE nuclear number between the blastocysts were analyzed using a general linear mixed model. An approximate t-test was used to compare differences in nuclei number (SAS 9.22 PROC MIXED, SAS Institute Inc, Cary, NC). Logistic regression analysis (SAS PROC LOGISTIC) was used to compare the total embryo numbers per stage per day on d3 and d5. Hatching/hatched to unhatched blastocysts, TE: ICM and abnormal nuclei:normal nuclei were compared through this same analytical approach. A P < 0.05 was considered significant. Deubiquitinating enzymes localizing in the ICM served as a point of reference for morphometric analysis.ResultsEquivalent numbers of embryos reached the 8C stage on d3 in both single and sequential embryo culture media. More d5 embryos reached the blastocyst stage, had greater numbers of normal nuclei, hatched and had significantly more trophoblast cells, but not ICM cells, when cultured in the single media compared to the sequential media.ConclusionSingle medium yields higher quality blastocyst embryos on d5 than sequential media. This supports the idea that single stage medium is more beneficial in human assisted reproduction procedures when compared to sequential media. ObjectiveDetermine effects of single versus sequential culture media on murine embryo quality and developmental attributes, which are not accessible in human embryos. Determine effects of single versus sequential culture media on murine embryo quality and developmental attributes, which are not accessible in human embryos. DesignMurine embryos cultured in commercially available single and sequential media were used to identify optimal preimplantation embryo quality and development attributes. Murine embryos cultured in commercially available single and sequential media were used to identify optimal preimplantation embryo quality and development attributes. Materials and MethodsMurine zygotes (n=40) were cultured in commercially available single or sequential media for 5 days. Images (200x, 400x) made using inverted phase contrast microscopes with differential contrast on developmental day 3 (d3) to facilitate quantification of the number of embryos developing to the 8-cell stage or greater in the two media. On d5, images were made and embryos fixed. Comparing and contrasting embryo images from the two types of media facilitated identification of an inner cell mass (ICM), nuclear quality assessment, and quantification of the total number of cells per embryo, ICM and trophoblast. The least square means (LSM) of total nuclei number, ICM nuclear number, and presumptive TE nuclear number between the blastocysts were analyzed using a general linear mixed model. An approximate t-test was used to compare differences in nuclei number (SAS 9.22 PROC MIXED, SAS Institute Inc, Cary, NC). Logistic regression analysis (SAS PROC LOGISTIC) was used to compare the total embryo numbers per stage per day on d3 and d5. Hatching/hatched to unhatched blastocysts, TE: ICM and abnormal nuclei:normal nuclei were compared through this same analytical approach. A P < 0.05 was considered significant. Deubiquitinating enzymes localizing in the ICM served as a point of reference for morphometric analysis. Murine zygotes (n=40) were cultured in commercially available single or sequential media for 5 days. Images (200x, 400x) made using inverted phase contrast microscopes with differential contrast on developmental day 3 (d3) to facilitate quantification of the number of embryos developing to the 8-cell stage or greater in the two media. On d5, images were made and embryos fixed. Comparing and contrasting embryo images from the two types of media facilitated identification of an inner cell mass (ICM), nuclear quality assessment, and quantification of the total number of cells per embryo, ICM and trophoblast. The least square means (LSM) of total nuclei number, ICM nuclear number, and presumptive TE nuclear number between the blastocysts were analyzed using a general linear mixed model. An approximate t-test was used to compare differences in nuclei number (SAS 9.22 PROC MIXED, SAS Institute Inc, Cary, NC). Logistic regression analysis (SAS PROC LOGISTIC) was used to compare the total embryo numbers per stage per day on d3 and d5. Hatching/hatched to unhatched blastocysts, TE: ICM and abnormal nuclei:normal nuclei were compared through this same analytical approach. A P < 0.05 was considered significant. Deubiquitinating enzymes localizing in the ICM served as a point of reference for morphometric analysis. ResultsEquivalent numbers of embryos reached the 8C stage on d3 in both single and sequential embryo culture media. More d5 embryos reached the blastocyst stage, had greater numbers of normal nuclei, hatched and had significantly more trophoblast cells, but not ICM cells, when cultured in the single media compared to the sequential media. Equivalent numbers of embryos reached the 8C stage on d3 in both single and sequential embryo culture media. More d5 embryos reached the blastocyst stage, had greater numbers of normal nuclei, hatched and had significantly more trophoblast cells, but not ICM cells, when cultured in the single media compared to the sequential media. ConclusionSingle medium yields higher quality blastocyst embryos on d5 than sequential media. This supports the idea that single stage medium is more beneficial in human assisted reproduction procedures when compared to sequential media. Single medium yields higher quality blastocyst embryos on d5 than sequential media. This supports the idea that single stage medium is more beneficial in human assisted reproduction procedures when compared to sequential media." @default.
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• W2013301740 title "Validating improved blastocyst quality and developmental attributes in single culture medium compare to sequential culture media using a murine model" @default.
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