Pericytes are essential to vascularization, but the purification and characterization of pericytes remain unclear. Smooth muscle actin alpha (alpha-SMA) is one marker [corrected] of pericytes. The aim of this study is to purify the alpha-SMA positive cells from bone marrow and study the characteristics of these cells and the interaction between alpha-SMA positive cells and endothelial cells. The bone marrow stromal cells were harvested from alpha-SMA-GFP transgenic mice, and the alpha-SMA-GFP positive cells were sorted by FACS. The proliferative characteristics and multilineage differentiation ability of the alpha-SMA-GFP positive cells were tested. A 3-D culture model was then applied to test their vascularization by loading alpha-SMA-GFP positive cells and endothelial cells on collagen-fibronectin gel. Results demonstrated that bone marrow stromal cells are mostly alpha-SMA-GFP positive cells which are pluripotent, and these cells expressed alpha-SMA during differentiation. The alpha-SMA-GFP positive cells could stimulate the endothelial cells to form tube-like structures and subsequently robust vascular networks in 3-D culture. In conclusion, the bone marrow derived pluripotent cells include [corrected] pericytes and can contribute to vascularization.