Purpose

To identify factors associated with isolation yields of ATP-binding cassette (ABC) superfamily member B5 (ABCB5)-positive limbal stem cells (LSCs) from human cadaveric donor eyes.

Methods

Whole eye globes were obtained from the Saving Sight eye bank, Kansas City, MO and the CorneaGen eye bank, Seattle, WA. ABCB5-positive LSCs were sorted by flow cytometry upon anti-ABCB5 monoclonal antibody staining within one week after donor death. The yields of live limbal epithelial cells in their entirety and of isolated pure ABCB5-positive LSC subsets were correlated with variables contained in the eye donors’ medical information.

Results

The mean isolation yield of live limbal epithelial cells and ABCB5-positive LSCs per donor eye was (340,000 ± 160,000 and 2,608 ± 1,842 respectively, mean ± SD). Stepwise regression analysis showed that cardiac disease-related death was the strongest negative predictor of the ABCB5-positive LSC isolation yield (p = 0.01). While we observed a trend for an age-related decline in the yield of ABCB5-positive LSCs, a statistically significant association could not be established (2% decrease/year, p = 0.11). Additionally, despite a trend for decreased isolation yields of total live limbal epithelial cells isolated from single donors with a longer time between death and tissue processing (p = 0.04), this did not affect the yields of purified ABCB5-positive LSC, which was independent of increasing time between death and tissue processing (p = 0.50).

Conclusions

Our study identifies cardiac disease-related death as a donor variable significantly associated with lower ABCB5-positive LSC isolation yields.

中文翻译：

---

调查与人类供体ABCB5阳性角膜缘干细胞分离产量相关的因素。

目的

为了确定与人尸体供体眼睛的ATP结合盒（ABC）超家族成员B5（ABCB5）阳性角膜缘干细胞（LSC）的分离产量相关的因素。

方法

整个眼球都是从密苏里州堪萨斯城的Saving Sight眼库和华盛顿州西雅图市的CorneaGen眼库获得的。供体死亡后一周内，通过抗ABCB5单克隆抗体染色，通过流式细胞术对ABCB5阳性LSC进行分类。完整的活的角膜缘上皮细胞的产量和分离的纯ABCB5阳性LSC亚群的产量与眼供体医学信息中包含的变量相关。

结果

每只供体眼睛的角膜缘活体上皮细胞和ABCB5阳性LSC的平均分离产率为（分别为340,000±160,000和2608±1,842，平均值±SD）。逐步回归分析表明，与心脏病相关的死亡是ABCB5阳性LSC分离产率的最强阴性预测因子（p  = 0.01）。尽管我们观察到了ABCB5阳性LSCs的产量与年龄相关的下降趋势，但仍未建立统计学上的显着关联（下降2％/年，p  = 0.11）。此外，尽管有从单个供体分离出的总活体角膜缘上皮细胞分离产量下降的趋势，而且死亡和组织加工之间的时间更长（p = 0.04），这不会影响纯化的ABCB5阳性LSC的产量，而该产量与死亡和组织加工之间的时间增加无关（p  = 0.50）。

结论

我们的研究确定与心脏病相关的死亡是与较低的ABCB5阳性LSC分离产率显着相关的供体变量。