Objective To explore the possibility of differentiating functional isletlike masses from bone marrow stromal cells induced by DMSO. Methods Adult Wistar rat bone marrow stromal cells (MSC) were isolated and cultured then induced for 3 days with serum free HDMEM media supplemented with 1% DMSO. The cells were then cultured in HDMEM supplemented with 10% FBS for 710 additional days. Morphological changes of the induced cells were observed under an inveted microscope. DTZ staining, immunocytochemical stainings were conducted to examine the production of insulin and glucagons. Glucose challenge and radio immunoassay were perfomed to evaluate the function of the aggregates, and RT-PCR was conducted to detect the expression of endocrine gene. Results While MSC cells were typically fibrocytelike, grapelike coherent clustered cells were observed in experiment group on the 3rd day after the inducement of DMSO, which were similar to pancreatic islets. DTZ staining of the cell aggregates was positive as the natural pancreatic islet. Immunocytochemistry also confirmed that these aggregates were positive for nestin on the first day after the inducement of DMSO, and positive for insulin, glucagen on the 10th day after the inducement; while the control group were negatively stained for the characteristics of endocrine. RT-PCR showed the insulin1, insulin2, glucagon and somatostaitin gene expression on the 10th day after the inducement of DMSO. Conclusion Isletlike