人眼角膜内皮细胞从血清基和合成储存介质保存的组织评估。

Human Corneal Endothelial Cell Assessment From Tissues Preserved in Serum-Based and Synthetic Storage Media.

机构信息

Institute of Ophthalmology, University College London, London, United Kingdom.

International Center for Ocular Physiopathology, The Veneto Eye Bank Foundation, Venice, Italy.

出版信息

Cornea. 2019 Nov;38(11):1438-1442. doi: 10.1097/ICO.0000000000002097.

DOI:10.1097/ICO.0000000000002097

PMID:31403530

Abstract

PURPOSE

To assess the difference between endothelial cells from tissues preserved in media supplemented with fetal bovine serum (FBS) and recombinant human serum albumin (rHSA).

METHODS

In a donor-matched study, 48 tissues were preserved for 28 days at 31°C in Cornea Max and Cornea Syn supplemented with FBS and rHSA, respectively. Endothelial cells were visualized by 2 masked observers before and after preservation. Endothelial cell density (ECD) and the number of iatrogenic folds were counted manually. Alizarin red staining and tight junction protein (Zonula Occludens-1) were used to assess cell morphology (hexagonality and polymorphism). Intraobserver and interobserver cell counts were recorded and analyzed. Wilcoxon and one-way analysis of variance tests were used, where P < 0.05 was deemed statistically significantly different.

RESULTS

Significant amount of iatrogenic folds were observed in the tissues supplemented with FBS compared with rHSA postpreservation (P = 0.0007). Approximately 69% and 71% hexagonal cells (P = 0.0303) and 29% and 26% polymorphic cells (P = 0.0234) were observed in the FBS and rHSA groups, respectively. Postpreservation, operator 1 counted 1766 cells/mm in FBS and 1864 cells/mm in rHSA. Operator 2 counted 1702 cells/mm in FBS and 1858 cells/mm in rHSA. ECD counts from FBS (interoperator) were statistically significant (P = 0.0429). However, significance was not observed in the ECD counts (interoperator) from the rHSA-preserved tissues (P = 0.8738).

CONCLUSIONS

rHSA-supplemented media allow better visualization of the corneal endothelial cells. This reduces the rate of discard observed due to counting errors. Use of rHSA improves the current standard of care and reduces the use of animal-derived products.

摘要

目的

评估在含有胎牛血清（FBS）和重组人血清白蛋白（rHSA）的培养基中保存的组织中的内皮细胞之间的差异。

方法

在一项供体匹配的研究中，将 48 个组织分别保存在 Cornea Max 和 Cornea Syn 中，其中分别添加 FBS 和 rHSA，在 31°C 下保存 28 天。在保存前后，由 2 位蒙面观察者观察内皮细胞。手动计数内皮细胞密度（ECD）和医源性褶皱数量。使用茜素红染色和紧密连接蛋白（Zonula Occludens-1）评估细胞形态（六边形和多态性）。记录并分析了观察者内和观察者间的细胞计数。使用 Wilcoxon 和单因素方差分析检验，P<0.05 被认为具有统计学显著性差异。

结果

与 rHSA 相比，保存后 FBS 组的组织中观察到大量医源性褶皱（P=0.0007）。在 FBS 和 rHSA 组中，分别观察到约 69%和 71%的六边形细胞（P=0.0303）和 29%和 26%的多态性细胞（P=0.0234）。保存后，操作者 1 在 FBS 中计数了 1766 个细胞/mm，在 rHSA 中计数了 1864 个细胞/mm。操作者 2 在 FBS 中计数了 1702 个细胞/mm，在 rHSA 中计数了 1858 个细胞/mm。FBS（观察者间）的 ECD 计数具有统计学意义（P=0.0429）。然而，rHSA 保存组织的 ECD 计数（观察者间）无统计学意义（P=0.8738）。