2023 - Full Presentation Schedule
Expression of both ubiquitin and histone proteins analyzed in blastocoel fluid from IVF embryos is correlated with positive implantation
Abstract
Introduction:
Due to an influx of patients seeking infertility treatments like IVF, the need to identify additional molecular markers, particularly proteins, in preimplantation embryos to predict successful implantation is essential. We propose assessing expression of proteins from the ubiquitin-proteasome family as well as histones in blastocoel fluid-conditioned media from IVF-embryos with known implantation outcomes. These data may provide evidence for embryologists to eventually predict implantation potential of an embryo with these assays.
Methods:
Blastocoel fluid conditioned media samples were collected from day-5-IVF embryos at the times of PGT-A biopsy. Media associated with embryos of poor morphology versus good morphology were used in this study. Total RNA and protein concentration were assessed with an Agilent Bioanalyzer. Blastocoel fluid-conditioned media samples were pooled and loaded onto a NuPAGE protein gel to detect total proteins with Colloidal Blue staining and then for Western Blots. Antibodies to detect ubiquitin and histones were used for the blots.
Results:
Preliminary results suggest that proteins can be detected in pooled blastocoel fluid conditioned media samples based on Colloidal Blue staining. The initial Western Blot assay detected some expression of ubiquitin.
Conclusions:
We hypothesize that the euploid grade AA embryos will present with stronger markers of histone and ubiquitin proteins as well as a higher concentration of protein in the blastocoel fluid, providing evidence that the embryos with stronger markers of said proteins will result in a successful implantation.
Expression of both ubiquitin and histone proteins analyzed in blastocoel fluid from IVF embryos is correlated with positive implantation
CASB 118 - Graduate Health Sciences
Introduction:
Due to an influx of patients seeking infertility treatments like IVF, the need to identify additional molecular markers, particularly proteins, in preimplantation embryos to predict successful implantation is essential. We propose assessing expression of proteins from the ubiquitin-proteasome family as well as histones in blastocoel fluid-conditioned media from IVF-embryos with known implantation outcomes. These data may provide evidence for embryologists to eventually predict implantation potential of an embryo with these assays.
Methods:
Blastocoel fluid conditioned media samples were collected from day-5-IVF embryos at the times of PGT-A biopsy. Media associated with embryos of poor morphology versus good morphology were used in this study. Total RNA and protein concentration were assessed with an Agilent Bioanalyzer. Blastocoel fluid-conditioned media samples were pooled and loaded onto a NuPAGE protein gel to detect total proteins with Colloidal Blue staining and then for Western Blots. Antibodies to detect ubiquitin and histones were used for the blots.
Results:
Preliminary results suggest that proteins can be detected in pooled blastocoel fluid conditioned media samples based on Colloidal Blue staining. The initial Western Blot assay detected some expression of ubiquitin.
Conclusions:
We hypothesize that the euploid grade AA embryos will present with stronger markers of histone and ubiquitin proteins as well as a higher concentration of protein in the blastocoel fluid, providing evidence that the embryos with stronger markers of said proteins will result in a successful implantation.