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在生物反应器中模拟体内培养条件有助于改善角膜保存中的组织质量。

In Vivo-Like Culture Conditions in a Bioreactor Facilitate Improved Tissue Quality in Corneal Storage.

机构信息

R. Schmid, Dr. A. Rossi, T. Schwarz, Dr. J. Hansmann, Translational Center Wuerzburg of the Fraunhofer Institute for Interfacial Engineering and Biotechnology (IGB), Röntgenring 11, 97070, Wuerzburg, Germany.

Dr. I.-S. Tarau, University Hospital Wuerzburg, Ophthalmic Clinic and Policlinic, Joseph-Schneider-Straße 11, 97080, Wuerzburg, Germany.

出版信息

Biotechnol J. 2018 Jan;13(1). doi: 10.1002/biot.201700344. Epub 2017 Sep 5.

Abstract

The cornea is the most-transplanted tissue worldwide. However, the availability and quality of grafts are limited due to the current methods of corneal storage. In this study, a dynamic bioreactor system is employed to enable the control of intraocular pressure and the culture at the air-liquid interface. Thereby, in vivo-like storage conditions are achieved. Different media combinations for endothelium and epithelium are tested in standard and dynamic conditions to enhance the viability of the tissue. In contrast to culture conditions used in eye banks, the combination of the bioreactor and biochrom medium 1 allows to preserve the corneal endothelium and the epithelium. Assessment of transparency, swelling, and the trans-epithelial-electrical-resistance (TEER) strengthens the impact of the in vivo-like tissue culture. For example, compared to corneas stored under static conditions, significantly lower optical densities and significantly higher TEER values were measured (p-value <0.05). Furthermore, healing of epithelial defects is enabled in the bioreactor, characterized by re-epithelialization and initiated stromal regeneration. Based on the obtained results, an easy-to-use 3D-printed bioreactor composed of only two parts was derived to translate the technology from the laboratory to the eye banks. This optimized bioreactor facilitates noninvasive microscopic monitoring. The improved storage conditions ameliorate the quality of corneal grafts and the storage time in the eye banks to increase availability and reduce re-grafting.

摘要

角膜是全球移植量最大的组织。然而,由于目前的角膜储存方法,供体的数量和质量受到限制。在这项研究中,采用了一种动态生物反应器系统来控制眼内压并在气液界面进行培养。从而实现了类似于体内的储存条件。在标准和动态条件下,测试了用于内皮细胞和上皮细胞的不同培养基组合,以提高组织的活力。与眼库中使用的培养条件相比,生物反应器和 biochrom 培养基 1 的组合可以保存角膜内皮细胞和上皮细胞。对透明度、肿胀和跨上皮电阻 (TEER) 的评估增强了类似于体内的组织培养的效果。例如,与静态储存条件下的角膜相比,测量到的光学密度明显更低,TEER 值明显更高(p 值<0.05)。此外,生物反应器中能够实现上皮缺损的愈合,表现为再上皮化和启动基质再生。基于获得的结果,衍生出一种易于使用的由两部分组成的 3D 打印生物反应器,将技术从实验室转化到眼库。这种优化的生物反应器便于进行非侵入性的微观监测。改进的储存条件改善了角膜移植物的质量和眼库中的储存时间,增加了可用性并减少了再次移植的需求。

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