Bone marrow-derived MSCs (BM-MSCs) is rare population represent about ~ 0.001–0.01% of bone marrow mononuclear cells. Because of their low abundance, in vitro culturing and expansion is essential requirement to obtain adequate numbers for research or clinical application. During long term culture, cells lose their proliferation, differentiation capacity and enter cellular senescence. In this study, we have tried to maximize number of cells separated from the same sample by allowing adhesion of bone marrow cells for 24h labeled with first adhesion cells (FACs) and re-culture of suspended cells again in a new flask labeled with re-culture of suspended cells (RSCs). We have characterized the morphology of FACs and RSCs. The results showed that the two types of cells have the same morphology. We concluded that we were able to isolate larger number of MSCs from the same sample