Adipose tissue is a rich source of mesenchymal stem cells (MSCs), which are multipotent cells capable of differentiating into various cell types, including osteocytes, adipocytes, and chondrocytes. Isolating stem cells from mouse adipose tissue, particularly from the inguinal fat pads, offers a valuable tool for stem cell research and regenerative medicine. This protocol outlines the method for isolating and culturing mouse adipose-derived stem cells (ADSCs) using collagenase digestion followed by centrifugation and resuspension in a culture medium supplemented with fetal bovine serum (FBS).