Some of in-vitro cell viability of islet of langerhans cell and hormone effects

Type1 Diabetes Mellitus is a chronic disease accompanied by the autoimmune destruction and loss of the beta cells of the pancreas related to some HLA alleles. Exogenous insulin therapy is the only treatment option to increase the quality of life and to prevent or to delay the chronic complications in patients with Type 1 diabetes. Intensive insulin therapy does not appear to achieve adequate success in preventing the chronic complications. ?n thetreatment of Type 1 diabetes, the studies directed to isolate the pancreatic islet cells and to perform the islet cell transplantation in to the liver via a lessin vasive way ( to the portal vein, by infusion) accelerated in 1990s. Hypoxia and the oxidative stres that developed during the isolation process may induce the cytokine-mediated apoptosis and necrosis and may lead to cell loss. There are data indicating that performing the culture of the islet cells before the transplantation may increase the volume of the cells in the preparations and prolongs the cell? s life. In our study, we aimed to demonstrate how the liveness of the cultured islet cells were affected after the irintroduction into the portal vein, to determine the effect of progesterone by adding progesterone to the islet hepatocyte co-culture under the in-vitro conditions and measuring the liveness of the islets by fluorescent staining and determining its effects on proliferation by counting the BrdU- stained isletsin flow cytometry, to perform the measurement of the function by glucose tolerance test and to determine the inter group differences.11860 male Wistar albino rats weighing 200-250 g, a ged from 2 to 5 months were used in the experiment. The examinations were performed at 0th and 48th hour in the 4 groups which were the islet culture group, the islet+ hepatocyte co-culture group, the Islet+ Progesteron culture group, Islet+ Hepatocyte+ Progesteron co-culturegroup. The hepatocyte isolation was based on the two-step isolation method of Seglen. The procedures for liveness and counting, were performed in Cauntes (Invitrogen-ABD). The fluorescent staining was performed with PI/FDA in order to measure the liveness of the islet cells. The cell proliferation was calculated in flow cytometry after the cells being marked by BrdU staining. ELIZA method was used for the glucose stimulation test. In the assessment of the liveness of the Islet cells, the highest increase between the 0th hourand 48th hour, was observed in the Islet+ Hepatocyte + Progesterone group. In the measurement of the islet cell proliferation, correspondingly to the liveness, the highest increase was observed in the Islet+Hepatocyte+ Progesteron group. While the results of glucose stimulation test revealedan increase in the insülin secretion at 48th hour in comparison with 0th hour, in the islet+ progesterone group in the presence of low glucose level, a decrease was observed in all groups in the presence of high glucose level. The data obtained were assessed by anova test and the results were found to be consistent with the findings.