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用于评估眼部通透性的角膜上皮细胞培养物的比较评价。

A comparative evaluation of corneal epithelial cell cultures for assessing ocular permeability.

作者信息

Becker Ulrich, Ehrhardt Carsten, Schneider Marc, Muys Leon, Gross Dorothea, Eschmann Klaus, Schaefer Ulrich F, Lehr Claus-Michael

机构信息

Saarland University, Biopharmaceutics and Pharmaceutical Technology, Saarbrücken, Germany.

出版信息

Altern Lab Anim. 2008 Feb;36(1):33-44. doi: 10.1177/026119290803600106.

Abstract

The purpose of this study was to evaluate the potential value of different epithelial cell culture systems as in vitro models for studying corneal permeability. Transformed human corneal epithelial (HCE-T) cells and Statens Serum Institut rabbit corneal (SIRC) cells were cultured on permeable filters. SkinEthic human corneal epithelium (S-HCE) and Clonetics human corneal epithelium (C-HCE) were received as ready-to-use systems. Excised rabbit corneas (ERCs) and human corneas (EHCs) were mounted in Ussing chambers, and used as references. Barrier properties were assessed by measuring transepithelial electrical resistance, and by determining the apparent permeability of markers with different physico-chemical properties, namely, fluorescein, sodium salt; propranolol hydrochloride; moxaverine hydrochloride; timolol hydrogenmaleate; and rhodamine 123. SIRC cells and the S-HCE failed to develop epithelial barrier properties, and hence were unable to distinguish between the permeation markers. Barrier function and the power to differentiate compound permeabilities were evident with HCE-T cells, and were even more pronounced in the case of C-HCE, corresponding very well with data from ERCs and EHCs. A net secretion of rhodamine 123 was not observed with any of the models, suggesting that P-glycoprotein or similar efflux systems have no significant effects on corneal permeability. Currently available corneal epithelial cell culture systems show differences in epithelial barrier function. Systems lacking functional cell-cell contacts are of limited value for assessing corneal permeability, and should be critically evaluated for other purposes.

摘要

本研究的目的是评估不同上皮细胞培养系统作为研究角膜通透性的体外模型的潜在价值。将转化的人角膜上皮(HCE-T)细胞和丹麦国家血清研究所兔角膜(SIRC)细胞培养在可渗透滤器上。SkinEthic人角膜上皮(S-HCE)和Clonetics人角膜上皮(C-HCE)作为即用型系统获得。将切除的兔角膜(ERC)和人角膜(EHC)安装在尤斯灌流小室中,并用作对照。通过测量跨上皮电阻以及测定具有不同物理化学性质的标志物(即荧光素钠盐、盐酸普萘洛尔、盐酸莫沙维林、氢溴酸噻吗洛尔和罗丹明123)的表观通透性来评估屏障特性。SIRC细胞和S-HCE未能形成上皮屏障特性,因此无法区分渗透标志物。HCE-T细胞具有屏障功能和区分化合物通透性的能力,在C-HCE中更为明显,与ERC和EHC的数据非常吻合。在任何模型中均未观察到罗丹明123的净分泌,这表明P-糖蛋白或类似的外排系统对角膜通透性没有显著影响。目前可用的角膜上皮细胞培养系统在上皮屏障功能方面存在差异。缺乏功能性细胞间接触的系统在评估角膜通透性方面价值有限,应针对其他目的进行严格评估。

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