Aims: c-KitPOS heart tissue-derived cardiac stem cells (CSCs) are helpful in myocardial infraction therapy but that the isolation and maintaining of heart tissue-derived CSCs are potential damage and complex.Whether the bone marrow deserves the seed cells alternative for heart tissue-derived CSCs is poorly defined.Notch signaling plays a critical role in the differentiation of stem cells and its role in c-KitPOS bone marrow-derived cardiac stem cells (c-KitPOS MCSC) are not well established.Methods and Results: After infected the Sprague Dawley rat bone marrow mesenchymal stem cells (BMSCs) with Notchl intracellular domain (NICD) adenovirus expression plasmid (NICD-Ad) for 8 days, the Notchl signaling was activated and cells underwent the myocardiocyte, smooth muscle cell and endothelial cell lineage differentiation.The c-KitPOS BMCSCs were successfully isolated from the BMSCs by using magnetic activated cell sorting (MACS) plus single cell cloning method.Among the four types of Notchl receptors, Notchl was predominantly expressed that tested by flow cytometry.Treated the c-KitPOS BMCSCs with Jagged1, the exogenous Notch1 ligand, led to an obviously activation of Notchl signaling and thus drove the cells to prominently differentiate into cardiomyocytes while much slight into smooth muscle cells and epithelial cells.However, the Notch1 inhibitor DAPT attenuated Jagged1-induced Notch1 activation and multi-lineage differentiation.Conclusions: There are c-KitPOS BMCSCs in the pool of BMSCs, suggesting an alternative seed cell for c-KitPOS heart tissue-derived CSCs.The activation of Notchl signaling contributes to the c-KitPOS BMCSCs multi-lineage differentiation, with prominent differentiation into cardiomyocytes, implying modulation of Notch1 signaling may be have potential application in the stem cell translational medicine.