Objective: Stem cell therapy is one of the most promising treatments for regenerative medicine. Curative therapy for diabetes mellitus mainly implies replacement of damaged insulin-producing pancreatic β cells with pancreas or islet-cell transplants. Due to the low number of donors, only a minority of patients benefit from this procedure. Pancreatic progenitor cells are a promising resource for regeneration of new islets. Therefore, the main aim of the current study was to isolate and characterize rat pancreatic stromal cells and to evaluate their potential to repair injured pancreatic islets. Materials and Methods: Male rats weighing approximately 200-250g were selected. Pancreas was exposed with PBS, removed and digested by collagenase P. Digestion terminated by cold PBS and tissues were washed two times, resuspended in DMEM supplemented with 10% serum, and were cultured on 6-wells plates. The cells were then subcultured after 5 days. RT-PCR was performed for the following genes: Nanog, Vimentin, Nucleostemin, Cyclin D1, Oct-4 and Sox2. Results: We successfully isolated islets and stromal cells from rat pancreas. Our results demonstrated that cultured stromal cells from rat pancreatic tissue express markers such as: Vimentin, Nucleostemin and Cyclin D1, which are known markers of the mesenchymal cells, but not the genes associated with Embryonic Stem Cells, such as Oct-4 and Sox2. Conclusion: Using a co-culture model in which damaged islets are grown on top of pancreatic stromal cells, we are now testing the capability of pancreatic stromal cells to repair damaged islets in rat model of diabetic. We are hopeful that our data help to find a new strategy for repairing the pancreatic islets of the diabetic patients in the future.