Differentiation Potential of Nestin (+) and Nestin (-) Cells Derived from Human Bone Marrow Mesenchymal Stem Cells into Functional Insulin Producing Cells

The feasibility of isolating and manipulating mesenchymal stem cells (MSCs) from human patients provides hope for curing numerous diseases and disorders. Recent phenotypic analysis has shown heterogeneity of MSCs. Nestin progenitor cell is a subpopulation within MSCs which plays a role in pancreas regeneration during embryogenesis. This study aimed to separate nestin cells from human bone marrow MSCs, and differentiate these cells into functional insulin producing cells (IPCs) compared with nestin cells. Manual magnetic separation was performed to obtain nestin cells from MSCs. Approximately 91±3.3% of nestin cells were positive for anti-nestin antibody. Pluripotent genes were overexpressed in nestin cells compared with nestin cells as revealed by quantitative real time-PCR (qRT-PCR). Following differentiation, flow cytometric analysis showed that 2.7±0.5% of differentiated nestin cells were positive for anti-insulin antibody in comparison with 0.08±0.02% of nestin cells. QRT-PCR showed higher expression of insulin and other endocrine genes in comparison with nestin cells. While immunofluorescence technique showed the presence of insulin and C-peptide granules in nestin cells. Therefore, our results introduced nestin cells as a pluripotent subpopulation within human MSCs which is capable to differentiate and produce functional IPCs.