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ARS Home » Plains Area » Clay Center, Nebraska » U.S. Meat Animal Research Center » Livestock Bio-Systems » Research » Publications at this Location » Publication #418209
Research Project: Improving Lifetime Productivity in Swine using Systems Biology and Precision Management Approaches

Location: Livestock Bio-Systems

Title: Isolation and characterization of extracellular vesicles from maternal and embryonic secretions during the initiation of porcine conceptus elongation

Author
item VYZOUREK, BENJAMIN - University Of Nebraska
item Miles, Jeremy
item Snider, Alexandria - Alex
item Rempel, Lea
item PANNIER, ANGELA - University Of Nebraska

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 9/15/2024
Publication Date: N/A
Citation: N/A

Interpretive Summary:

Technical Abstract: Improving the reproductive efficiency of pigs is crucial to meet the escalating global demands for food supply sustainability. Porcine pregnancy success is heavily influenced by a critical period of prenatal development between days 9 to 12 of gestation, when the embryo rapidly elongates from a spherical blastocyst (~1mm) to a long filamentous morphology (>100mm), which requires extensive signaling between maternal and embryonic tissues. Extracellular vesicles (EVs) containing proteins, mRNAs, and miRNAs have been detected in abundance in reproductive fluids, providing evidence for their role in cell-to-cell communication in mammalian reproduction. Specifically, known signaling pathways and interactions between tissues suggest miRNAs carried in EVs may be integral to regulating developmental processes including the initiation of embryo elongation. To characterize EVs at the initiation of elongation, gilts were bred and harvested at days 9, 10, or 11 of gestation. Reproductive tracts were removed, and each uterine horn was flushed with RPMI-1640 medium. EVs were isolated from the flushing material using ultracentrifugation, tangential flow filtration, and ultrafiltration. This size-based approach yielded two populations enriched in small or large EVs, allowing for the characterization of relevant subpopulations. All particles were analyzed via nano-flow cytometry. Analysis revealed that small EVs exhibited lipid-positive percentages of 84 ± 14, 86 ± 10, and 79 ± 13% on days 9, 10, and 11, respectively, while large EVs were 91 ± 6, 88 ± 9, and 80 ± 12% lipid-positive on those days, respectively. Small EVs averaged 69 ± 2, 65 ± 3, and 64 ± 3nm in diameter, and large EVs averaged 200 ± 74, 191 ± 57, and 206 ± 40nm on days 9, 10, and 11, respectively. In conclusion, these optimized isolation techniques, combined with nano-flow cytometry analysis, ensure pure EV populations for future downstream RNA sequencing analyses. USDA is an equal opportunity provider and employer.