Young porcine endocrine pancreatic islets cultured in fibrin show improved resistance toward hydrogen peroxide
Young porcine endocrine pancreatic islets cultured in fibrin show improved resistance toward hydrogen peroxide
复制标题
在纤维蛋白中培养的幼猪内分泌胰岛显示出对过氧化氢的抵抗力得到改善
作者：
C. Kuehn;J. Lakey;Morgan W. Lamb;P. Vermette
目的：研究纤维蛋白支架对双氧水包埋的幼年猪内分泌胰岛的保护作用。方法：分离和体外成熟后，将200组猪胰岛当量（IEQ）幼猪包埋在200µL纤维蛋白凝胶中，并暴露于2种浓度（10和100µM）的过氧化氢（H2O2）中，研究纤维蛋白对胰岛细胞凋亡刺激的保护能力。作为对照，将幼猪胰岛接种于组织培养聚苯乙烯（TCPS）孔板中，并暴露于相同浓度的H2O2中。然后研究胰岛的完整性、活力和功能。结果：在形态学上，与在TCPS板中培养的胰岛相比，纤维蛋白凝胶包埋的胰岛在暴露于H2O2时能更好地保存其完整性。免疫荧光染色显示胰岛素和胰高血糖素在纤维蛋白培养的胰岛中表达较高。总的来说，H2O2孵育导致胰岛素和胰高血糖素表达降低。TUNEL实验显示，与包埋在纤维蛋白中的胰岛细胞相比，在TCPS板中培养的胰岛细胞凋亡数量增加。在TCPS培养板中培养并暴露于H2O2的胰岛对葡萄糖刺激分泌胰岛素的能力下降，而嵌入纤维蛋白的胰岛则保持对葡萄糖的反应。提取并定量捕获纤维蛋白中的胰岛素，揭示提取液中的胰岛素。结论/解释：纤维蛋白对暴露于过氧化氢的幼猪内分泌胰岛具有保护作用。
Aim: To study the protective effect of a fibrin scaffold toward embedded young porcine endocrine pancreatic islets from hydrogen peroxide within the context of islet encapsulation in transplantation. Methods: After isolation and in vitro maturation, groups of 200 young porcine islet equivalents (IEQ) were embedded in a 200 µL fibrin gel and exposed to 2 concentrations (10 and 100 µM) of hydrogen peroxide (H2O2) to investigate the ability of fibrin to protect islets against apoptotic stimuli. As a control, young porcine islets were seeded in tissue culture polystyrene (TCPS) well plates and exposed to the same H2O2 concentrations. Islet integrity, viability and function were then investigated. Results: Morphologically, the integrity of islets embedded in fibrin gels was better preserved compared with that of islets cultured in TCPS plates, when exposed to H2O2. Immunofluorescence staining showed that insulin and glucagon expression was higher in islets cultured in fibrin. Overall, H2O2 incubation led to decreased insulin and glucagon expression. A TUNEL assay revealed elevated numbers of apoptotic cells for islets cultured in TCPS plates when compared with those embedded in fibrin. Islets cultured in TCPS plates and exposed to H2O2 had diminished ability to secrete insulin in response to glucose stimulation, whereas islets embedded in fibrin maintained their glucose responsiveness. Insulin trapped in fibrin was extracted and quantified, revealing insulin in the extract. Conclusions/Interpretation: Fibrin has a protective effect on young porcine endocrine pancreatic islets exposed to hydrogen peroxide.
影响因子：
4.1
作者：
Weber, Laney M.;Hayda, Kirsten N.;Anseth, Kristi S.
通讯作者：
Anseth, Kristi S.