Bone marrow increases human islets insulin positive cells in co-culture: quantification with flow cytometry

Abstract

We have previously demonstrated that allogeneic human bone marrow (BM) supports human islet function and longevity in vitro. We hypothesize that BM supporting human islets may include to increase β-cell in cultured islets. In this study, we developed a method to quantify insulin-producing β cells from cultured islets by using immunofluorescent staining and flow cytometry analysis to explore this possibility. The results show that human islets cocultured with BM for 39 days contained a significantly higher number of insulin-positive β cells (42.3% ± 4.5%) compared to the islet-only cultures (1.15% ± 0.78%), and increased insulin release levels evaluated by ELISA is consistent with increased β cells in same culture condition. Human islet culture with BM significantly increase β-cells while islet only culture lost β-cells in same culture period supports the possibility of BM increasing β-cells in cultured islets.

Anti-apoptotic Effects of Bone Marrow on Human Islets: A Preliminary Report

“…In our study, bone marrow cells blocked the initiation of cell apoptosis by inhibiting P2X7R activation, protecting human islet β-cell from apoptosis ( Figure 6 ). Furthermore, islet function in terms of insulin release was maintained in comparison with the deterioration occurred in islet only group ( Figure 5A ), which is consistent with our previous studies [ 38 ]. The correlation of insulin and IL-1β release demonstrated that the reduction of insulin concentration might be related with the increasing release of IL-1β in islet only cultures, while BM co-culture prevented the increase in IL-1β.…”