An Advanced Murine Model for Nonalcoholic Steatohepatitis in Association with Type 2 Diabetes

This method can help answer questions in the immunometabolism field about the contribution of immune cells to obesity-induced inflammation in adipose and steatohepatitis in hepatic tissue. These techniques have been optimized for the isolation of adipose and hepatic immune cells in a diet-induced model of nonalcoholic steatohepatitis achieved through a non-specific pathogen-free rodent housing. These methods can help to deepen our understanding of the immunological mechanisms involved in the maintenance of metabolic homeostasis.

Generally, individuals new to this technique will struggle with maximizing the yield of functionally viable dissociated cells and setting up the proper gating in flow cytometry. To generate an adipose tissue single-cell suspension, spray the chest of a euthanized mouse with 70%ethanol, and carefully make a five to six-centimeter central incision through the integument and abdominal wall along the entire length of the rib cage to expose the pleural cavity and heart. Using a 26-gauge needle, inject at least 10 milliliters of 0.9%saline solution into the apex of the left ventricle, and use scissors to open the peritoneal cavity.