Mesenchymal stem cells (MSCs) are multipotent stem cells – ability to differentiate into several cell types. Previous studies have showed a high number of these cells can be found in adipose tissue. This study has established a simple protocol for isolation and culture of MSCs from mouse adipose tissue using minimal requirements. With non-enzymatic approach, we successfully isolated and cultured the MSCs. Isolated MSCs were identified through morphology and ability of differentiation into adipocyte-like cells under the presense of stimuli in the culture medium.