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Young porcine endocrine pancreatic islets cultured in fibrin show improved resistance toward hydrogen peroxide

机译:用纤维蛋白培养的年轻猪内分泌胰腺胰岛对过氧化氢的抵抗力增强

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摘要

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.
机译:目的:研究在移植胰岛包封的情况下,纤维蛋白支架对过氧化氢埋入的年轻猪内分泌胰岛的保护作用。方法:分离并体外成熟后,将200个年轻猪胰岛当量(IEQ)组嵌入200 µL纤维蛋白凝胶中,并暴露于2种浓度(10和100 µM)的过氧化氢(H2O2)中,以研究纤维蛋白的能力。保护胰岛免受凋亡刺激。作为对照,将年轻的猪胰岛接种到组织培养聚苯乙烯(TCPS)孔板上,并暴露于相同的H2O2浓度。然后研究胰岛的完整性,生存力和功能。结果:从形态上讲,当暴露于H2O2时,与在TCPS平板中培养的胰岛相比,包埋在纤维蛋白凝胶中的胰岛的完整性得到了更好的保存。免疫荧光染色显示,在纤维蛋白中培养的胰岛中,胰岛素和胰高血糖素的表达较高。总体而言,H2O2孵育导致胰岛素和胰高血糖素表达降低。 TUNEL分析显示,与包埋在纤维蛋白中的胰岛相比,TCPS平板中培养的胰岛的凋亡细胞数量增加。在TCPS平板中培养并暴露于H2O2的胰岛响应葡萄糖刺激而分泌胰岛素的能力降低,而包埋在纤维蛋白中的胰岛则保持了其葡萄糖响应性。提取和定量捕获在纤维蛋白中的胰岛素,从而揭示提取物中的胰岛素。结论/解释:纤维蛋白对接触过氧化氢的年轻猪内分泌胰岛具有保护作用。

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