Bone Marrow Mesenchymal Stem Cells (BMMSCs) are adult stem cells with the capacity for selfrenewal and multilineage differentiation. Endothelial Progenitor Cells (EPCs) play a fundamental role in post-natal vascular repair. Both BMMSCs and EPCs are rare in bone marrow and therefore optimizing isolation and culture is required before they can be applied as parts of scientific research and clinical therapies. The aim of this study was to develop an efficient method to isolate and expand both BMMSCs and EPCs in the same rat bone marrow at the same time. In this study, rat BMMSCs and EPCs were isolated by differential adhesion method and cultured in vitro. Four passaged BMMSCs were CD29+, CD90+ and CD45-. EPCs were CD34+ and FLK-1+. BMMSCs differentiate in vitro into chondrogenic and adipogenic cells based on the positive staining with Alcian Blue, Type 2 Collagen and Oil Red O. In conclusion, the differential adhesion method was found to be a convenient and reliable method for the isolation and culture of both BMMSCs and EPCs from one bone marrow donor sample at one time.