Mesenchymal stem cells (MSCs) are multipotent progenitor cells that can differentiate into a variety of cell types, including adipocytes, osteocytes, and chondrocytes. Adipose tissue is a rich and easily accessible source of these stem cells, known as adipose-derived stem cells (ADSCs). These cells have gained significant interest in regenerative medicine due to their ability to differentiate into various tissues, their high proliferative capacity, and their immunomodulatory properties. Additionally, ADSCs can be isolated with relative ease from small tissue samples, such as inguinal fat pads, making them an ideal model for cell-based therapies and tissue engineering applications. The protocol outlined here describes a method for isolating mouse adipose-derived stem cells (ADSCs) from inguinal fat tissue using collagenase digestion, followed by centrifugation and resuspension in a culture medium. This process allows for the efficient isolation of viable ADSCs, which can be further expanded or differentiated for experimental purposes. The simplicity and reproducibility of this protocol make it suitable for both basic research and applications in stem cell therapy.