Semi-Automated Isolation of the Stromal Vascular Fraction from Murine White Adipose Tissue Using a Tissue Dissociator

We are interested in brown and beige subtypes of adipocyte which dissipate energy in the form of heat, while white adipocyte generally store energy as Our goal is to develop a new treatment for obesity and diabetes based on promotion of energy expenditure, rather than suppression of energy intake. We have identified a transcription factor, NFI, as a critical regulator of brown adipocyte differentiation by a genome-wide open chromatin analysis. NFI and the master transcriptional regulator of adipogenesis, PPAR-gamma, co-localize as the brown-fat-specific enhancers.

In the process of isolating stromal vascular fraction from mouse adipose tissue, manual mincing and collagenase digestion of adipose tissue can lead to experimental variability and contamination. Our method uses a tissue dissociator for collagenase digestion, which facilitates SVF isolation, reduce experimental variability and contamination, and improves reproducibility. Experiment using individually-differentiated adipocyte are indispensable for research in our field.

Our protocol would offer easy and reproducible SVF isolation for subsequent adipocyte differentiation and would accelerate research in our community. Currently, we are investigating the role of NFI of all-body metabolism, such as body weight and glucose, using gain and loss of function mouse models. We will also explore other pharmacological activation of NFI for each downstream pathway in near future.