Mesenchymal stem cells (MSCs) are characterized by their ability of self-renewal paired with the capacity to differentiate into multiple mesenchymal cell lineages. Numerous studies have reported beneficial effects of MSCs in tissue repair and regeneration. After in vivo administration, MSCs home to and engraft to injured tissues. However, the molecular mechanisms are not clear. Osteopontin (OPN) has been found to be elevated in response to injury and inflammation and its role on cell mobilization has been studied. Therefore, the facts imply that OPN may contribute to the recruitment of MSCs to the sites of injury. In this study, using transwell assay, we found that rat bone marrow-derived mesenchymal stem cells (rMSCs) migrated towards OPN in a concentration-dependent manner. To further examine the involved molecular mechanisms for OPN-induced rMSCs migration, RT-PCR, and Western blot were used to detect the expressions of integrin β1 and CD44v6, the two receptors of OPN. OPN promoted integrin β1 mRNA and protein expression while CD44v6 mRNA level was not altered. Blockade of integrin β1 also inhibited OPN-induced rMSCs migration, indicating the possible involvement of integrin β1 in OPN-induced migration in rMSCs. Our data have shown for the first time that OPN increases integrin β1 expression in rMSCs and promotes rMSCs migration through the ligation to integrin β1.