Project description:Comparison of bovine embryo transcriptome at 16-cell (96 hpi) and blastocysts (192 hpi) stages in presence of co-culture systems (BOEC or VERO) in SOF medium
Project description:Serum is commonly added to culture medium used for producing bovine embryos in vitro. Here it was tested whether addition of bovine serum to embryo culture medium increases the percent of embryos becoming blastocysts and alters blastocyst gene expression in a manner that could affect competence to establish pregnancy or that could lead to dysregulated fetal development. A secondary objective was to test whether actions of serum depend upon the specific medium used for embryo culture. Media used were synthetic oviduct fluid medium bovine embryo 2 (SOF-BE2) and a commercial medium termed BO-IVC. Fetal bovine serum and adult bovine serum were added at a final concentration of 10% (v/v) at either day 1 or 5 of development. Overall, addition of serum to culture medium increased blastocyst production. Effects of fetal bovine serum added at day 5 of development caused moderate changes in gene expression in the resulting blastocysts, with differentially expressed genes being those with an adjusted p-value of < 0.01 and an absolute log2 fold change > |1|. Serum resulted in 114 differentially-expressed genes for embryos cultured in SOF-BE2 and 108 embryos cultured in BO-IVC. Only 15 genes were regulated by serum similarly for both media including transcription factors ASCL2 (downregulated) and ZSCAN4 (upregulated) and the phosphoserine aminotransferase PSAT1 that has been implicated in fetal growth in mice. Genes of the melanoma antigen gene family that regulate degradation of specific transcription factors were also upregulated by serum. Expression data on eight biomarker genes were also used to calculate an embryo competence index that has been shown to predict embryo ability for survival after embryo transfer. Serum lowered the embryo competence index for both media, suggesting serum could compromise embryonic survival. In conclusion, actions of serum on the preimplantation embryo include changes in gene expression indicative of reduced embryo competence. Some changes in gene expression could potentially lead to alterations in fetal development
Project description:To analyze impact of co-culture presence and type of feeder cell lines, we compared the transcriptome of bovine blastocyst (192 hpi) using a new custom bovine microarray.
Project description:Comparison of gene expression from expanded bovine blastocysts collected 7 days after fertilization and produced in vivo vs in vitro-SOF-OPU
Project description:EtOH-Lipid form serum have been added in BSA-supplemented SOF during post-compaction development of IVP bovine embryos. Lipid-treated blastocysts were compared with control treated blastocysts (no lipid addition to BSA, EtOH balanced).
Project description:Comparison of gene expression from expanded bovine blastocysts collected 7 days after fertilization and produced in vivo vs in vitro-SOF-OPU Two kinds of 7 days post fertilization bovine embryos, in-vivo 7 days blastocysts vs. In vitro 7 days blastocysts, Biological replicates: 4 in-vitro, 4 in-vivo, protocol,extract and semen shared. Dye swap.
Project description:EtOH-Lipid form serum have been added in BSA-supplemented SOF during post-compaction development of IVP bovine embryos. Lipid-treated blastocysts were compared with control treated blastocysts (no lipid addition to BSA, EtOH balanced). Amplified anti-sense RNA from each sample (4 pools of control and 4 pools of treated blastocysts) were labeled then hybridized on oligonucleotide microarrays following a dye-swap design (ex: pool 1 ctl in red vs. pool 1 treatment in green and then pool 1 ctl in green vs. pool 1 treatment in red).
Project description:To analyze impact of co-culture presence and type of feeder cell lines, we compared the transcriptome of bovine 16-cell (96 hpi) using a new custom bovine microarray.
Project description:In this study, we focused on the changes of mRNA expression in bovine oviduct epithelial cells in vitro co-cultured with embryos during preimplantation development in order to identify genes, which might be involved in embryo-maternal communication. For this purpose, we used large-scale cDNA microarray hybridizations to identify the genes differentially regulated in bovine oviduct epithelial cells (Boec) during in vitro culture. The main objective was to identify genes which are differentially expressed due to the presence or absence of bovine embryos on cell monolayers. Global transcriptional profiling was performed using 13 days Bovine oviduct epithelial cells (Boec) cultured in vitro with SOF media RNA as control samples (BOEC_SOF_CTL, n=3) for comparison to the experimental samples taken at the same culture time (day 13) and stimulated by the presence of bovine embryos during the last 8 days of culture (BOEC_SOF+EMB, n=3).Gene expression analysis was carried out between Boec with or without stimulation of embryos representing a total of 6 slides (dye swap protocol).
