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Abstract Diabetes constitutes a worldwide epidemic that affects all ethnic groups. Cell therapy is one of the best alternatives for treatment. Nestin is a marker for neural stem cells and has been reported in regenerating pancreas. It was suggested that it plays a pivotal role as an intermediate regulator governing the differentiation of stem cells in the process of their differentiation into insulin-secreting cells. In this study, we isolated nestin-positive mesenchymal stem cells from adult human bone marrow to evaluate their efficiency to differentiate into insulin-producing cells as compared with nestin negative fraction. Nestin rich and nestin poor cells were isolated from HBM-MSCs by magnetic cell separation, following their expansion, differentiation was performed Trichostatin-A/GLP-1 protocol. At the end of differentiation, cells were evaluated by expression of pancreatic endocrine genes, immunofluorescence using the confocal microscopy, flow cytometry as well as human insulin and c-peptide release in response to increasing glucose concentrations. Confocal microscopy analysis showed 91.1% nestin-positive cells in the rich population while 1.28% in the poor one. At the end of differentiation, the relevant endocrine genes including INS, GCG and SST were expressed significantly higher in nestin rich cells than in nestin poor cells. The percentages of generated IPCs detected by flow cytometry and immunofluorescence were significantly higher in nestin rich cells than in nestin poor cells and this was the case after applying ELISA test for determination of insulin and c-peptide release after exposure to different concentrations of glucose. Thus, our findings present evidence that nestin-positive cells are a subpopulation of MSCs capable of generating IPCs in a higher percentage than its negative counterpart. |