Milk-Derived EVs from Different Animal Sources: An Overview on Their Detection, Isolation and Pleiotropic Exerted...

Area of Science:

• Biochemistry and Molecular Biology
• Animal Science
• Biotechnology

Background:

• Milk is a nutrient-rich matrix essential for growth, containing milk-derived extracellular vesicles (mEVs).
• These mEVs, a type of food-derived EV (fEV), have potential therapeutic applications due to their biocompatibility and stability.
• Challenges exist in standardizing mEV isolation and characterizing their molecular cargo.

Purpose of the Study:

• To provide a comparative analysis of mEVs across various mammalian species.
• To discuss critical isolation techniques to minimize contamination.
• To review current and advanced detection methods, particularly flow cytometry.

Main Methods:

• Comparative analysis of mEVs from bovines, buffaloes, yaks, camels, goats, pigs, horses, donkeys, and humans.
• Discussion of isolation protocols focusing on minimizing milk fat globule and casein micelle contamination.
• Review of conventional and imaging flow cytometry (FC) techniques, including bead-based assays and generic EV markers.

Main Results:

• mEVs exhibit distinct functional signatures across different mammalian species.
• Effective isolation requires minimizing interference from milk fat globules and casein micelles.
• Advanced FC techniques, including IFC, offer improved detection, but require careful selection of markers and antibody clones for accuracy.

Conclusions:

• Standardized isolation and detection methods are crucial for reproducible mEV research.
• Flow cytometry, especially IFC, holds promise for characterizing mEVs across species.
• Understanding inter-species mEV variability is key for leveraging their nutritional and therapeutic potential.