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富含全氟癸烷的纤维蛋白基质用于人胰岛培养。

Perfluorodecalin-enriched fibrin matrix for human islet culture.

机构信息

Islet Transplant Research Group, Nuffield Department of Surgical Sciences, University of Oxford, Oxford, UK.

出版信息

Biomaterials. 2011 Dec;32(35):9282-9. doi: 10.1016/j.biomaterials.2011.08.044. Epub 2011 Sep 6.

DOI:10.1016/j.biomaterials.2011.08.044
PMID:21899883
Abstract

Disruption of microenvironment and decrease in oxygen supply during isolation and culture lead to pancreatic islet injury and their poor survival after transplantation. This study aimed to create a matrix for culturing islets, using fibrin as scaffold and perfluorodecalin as oxygen diffusion enhancing medium. Human pancreatic islets were divided in four groups: control, islets cultured in fibrin, islets in fibrin containing non-emulsified perfluorodecalin, and finally islets in fibrin supplemented with emulsified perfluorodecalin. After an overnight culture, cell damage (viability, proinsulin and insulin unregulated release, apoptosis (caspase-3 activation), secretory function, and presence of hypoxia markers (HIF-1a and VEGF expression) were assessed. Islets cultured in a matrix, had similar islet viability to controls (no matrix) but decreased levels of active caspase-3 and unregulated hormone release, but high level of hypoxia markers expression. Although the supplementation of fibrin with non-emulsified perfluorodecalin improves secretory response, there was no decrease in hypoxia markers expression. In contrast, emulsified perfluorodecalin added to the matrix improved islet function, islet viability and maintained level of hypoxia markers similar to control. Fibrin matrix supplemented with emulsified perfluorodecalin can provide a beneficial physical and chemical environment for improved pancreatic human islet function and viability in vitro.

摘要

在隔离和培养过程中,微环境的破坏和氧气供应的减少会导致胰岛损伤,并且它们在移植后生存状况不佳。本研究旨在创建一种用于培养胰岛的基质,使用纤维蛋白作为支架,全氟癸烷作为氧气扩散增强介质。将人胰岛分为四组:对照组、纤维蛋白培养的胰岛、含非乳化全氟癸烷的纤维蛋白中的胰岛,以及最后一组是纤维蛋白中添加乳化全氟癸烷的胰岛。经过一夜的培养,评估了细胞损伤(活力、胰岛素原和胰岛素非调节性释放、细胞凋亡(半胱天冬酶-3 激活)、分泌功能以及缺氧标志物(HIF-1a 和 VEGF 表达)的存在情况。在基质中培养的胰岛与对照组(无基质)具有相似的胰岛活力,但活性半胱天冬酶-3 和非调节性激素释放水平降低,但缺氧标志物的表达水平较高。虽然向纤维蛋白中添加非乳化全氟癸烷可以改善分泌反应,但缺氧标志物的表达并没有降低。相比之下,向基质中添加乳化全氟癸烷可以改善胰岛功能、胰岛活力,并使缺氧标志物的表达水平与对照组相似。补充乳化全氟癸烷的纤维蛋白基质可以为体外改善人胰岛功能和活力提供有益的物理和化学环境。

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