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.
Highlights
Diabetes is a condition whereby the body is not able to regulate levels of glucose in the blood, resulting in too much glucose being present in the blood
Pancreatic islet transplantation is a promising treatment for patients with Type 1 diabetes mellitus, which results in hyperglycemia due to islet β cells damage [1,2,3,4]
In evaluation of bone marrow (BM) co-culture human islet during different culture days using same method, we found that β cell population from islet with BM co-culture significantly increases from 14 to 39 day in cultures (28.15% ± 3.4% on day 14 and 42.3% ± 4.5% on day 39 cultures) (Figure 4(a)) and function evaluation found insulin release increase but too much variation to statistic sig
Summary
Diabetes is a condition whereby the body is not able to regulate levels of glucose (a sugar) in the blood, resulting in too much glucose being present in the blood. We did not successfully detect positive FluoZin-3 in dissociated monolayer islet cells in long term culture in vitro because this method stained some kind bone marrow cell like monocyte [19] and T cell [20]. This method will result false positive β-cells in islet and bone marrow co-culture. Wang et al / Journal of Diabetes Mellitus 1 (2011) 109-117 to quantify insulin positive cells from a culture containing a heterogeneous mixture of islet cells Using this technique, we were able to quantitate insulin-producing β cells from culture islets with or without BM. Our studies support the hypothesis that BM supporting human islet β cell function includes increase β cell population while not in islet only culture
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