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: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: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: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).
Project description:Retinoic acid receptor gamma (RARγ) plays a critical but poorly understood role in early embryo development and stem cell pluripotency. Here, we investigated the effects of the RARγ-specific inhibitor LY2955303 (LY) on in vitro-produced bovine embryos. Treatment with LY significantly increased blastocyst rates and quality, demonstrating its potential to enhance IVF outcomes in cattle. Single-embryo RNA sequencing using Smart-seq indicated that LY promotes metabolic reprogramming (upregulating glycolysis and TCA cycle activity) while suppressing apoptosis and inflammatory responses. LY- treatment starting from 16-cell stage led to enhanced glycolysis in blastocysts compared with exposure from 2-cell stage. Furthermore, LY-treatment also upregulated oxidative phosphorylation in bovine embryonic stem cells (ESCs). These findings establish RARγ as an important regulator of bovine embryo development, with its inhibition offering a novel strategy to optimize IVF embryo culture systems for livestock production.
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:Research in JCPyV infection of primary astrocytes is limited. However, viral infection in a transformed glial cell line (SVGAs), mostly composed of astrocytes, is well studied. RNA sequencing was performed at 24 hours post infection (hpi), 48 hpi, and 96 hpi. Timepoints established to be important in both cell types in the JCPyV infectious cycle.
