Characterization of Adipocyte-Derived Extracellular Vesicle Secretion Using a CD63-GFP Reporter Mouse Model In Vivo and In Vitro

We study adipocyte-derived extracellular vesicle, EVs, to define their roles in adipose tissue, particularly how they regulate the crosstalk between mature adipocytes and their progenitor cells. There are currently no in vivo systems to study adipocyte EV secretions, and the existing purification protocols are not yet fully optimized. To begin, use sterile technique to excise inguinal adipose tissue from euthanized mice.

Place one gram of tissue into a two milliliter tube containing 0.5 milliliters of digestion buffer. Using sterile scissors, mince the tissue into pieces no larger than one millimeter. Suspend the minced tissue in 10 milliliters of digestion buffer containing 0.5 milligrams per milliliter Liberase and 50 units per milliliter DNase I.Incubate the sample at 37 degrees Celsius on an orbital shaker.