Three-Dimensional Culture of Vascularized Thermogenic Adipose Tissue from Microvascular Fragments

This protocol describes the generation of three-dimensional vascularized and functional thermogenic fat from microvascular fragments, or MVFs. Current strategies for engineering thermogenic adipose tissue are complex and don't fully depict its multicellular and functional properties. MVFs are a single source of cells that enable vascularization and adipose tissue formation.

Being a simple sole-source method to create biological imitations of beige fat, MVFs hold significant potential for promoting the understanding or development of treatments for obesity and metabolic disease. Place the appropriately shaved and euthanized animal treated with 70%ethanol in a supine position and begin isolation of the inguinal fat. Lift the skin below the penis with a pair of scissors and perform the incision, starting at the center and cutting laterally, forming a V shape, before looping to the animal's backside to access the entire fat deposit.