Location: Reproduction Research
Project Number: 3040-31000-095-07-R
Project Type: Reimbursable Cooperative Agreement
Start Date: Apr 1, 2017
End Date: Feb 28, 2021
Specific Objective 1) Identify extraembryonic and embryonic gene transcripts and metabolites that regulate the initiation of pig embryo elongation in vivo and in vitro within a 3-D culture system. Specific Objective 2) Investigate the role of specific molecular factors and cellular components on the initiation of elongation within a 3-D culture system.
The overall strategy for Objective #1 is to utilize a global approach to identify critical molecular factors, both within the embryo and the uterine/culture environment, that regulate the initiation of pig embryo elongation. Embryos will be collected and encapsulated within a supporting hydrogel matrix, or collected in vivo at various time points. Those embryos identified as initiating elongation, as well as those that have failed to initiate, will be analyzed for alterations in critical molecular factors. For identifying factors within the embryo, a transcriptomic approach (i.e., RNA-seq) will be employed. For identifying soluble factors within the uterine/culture environment, a metabolomics approach (i.e., gas chromatography-mass spectrometry and ultra performance liquid chromatography-mass spectrometry) will be utilized. The overall strategy for Objective #2 is to evaluate the role of specifically identified critical factors and cellular components (i.e., uterine endometrial cells) on the initiation of pig embryo elongation, using the established 3-D culture. Factors identified from the literature that are hypothesized to play a role on elongation, will be incorporated into the culture system, either within the media, within the supporting hydrogel matrix, or via co-cultures. Encapsulated embryos will be monitored for their ability to initiate elongation, monitored through survival, morphological changes, cellular proliferation and differential gene expression for steroidogenesis, cellular proliferation and differentiation as well as estradiol production.