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长时间低温保存后培养的人角膜内皮细胞的改善

Improvement of human corneal endothelium in culture after prolonged hypothermic storage.

作者信息

Camposampiero D, Tiso R, Zanetti E, Ruzza A, Bruni A, Ponzin D

机构信息

Veneto Eye Bank Foundation, Venezia Mestre, Italy.

出版信息

Eur J Ophthalmol. 2003 Nov-Dec;13(9-10):745-51. doi: 10.1177/1120672103013009-1002.

DOI:10.1177/1120672103013009-1002
PMID:14700093
Abstract

PURPOSE

To evaluate whether the organ culture method for human cornea preservation may be applied to corneas stored for several days at 4 degrees C.

METHODS

The cell density, viability, and morphology of corneal endothelium were examined in 140 human corneas stored at 4 degrees C for the minimal time required for transport to the bank and for the preliminary controls of cornea status (1.6 +/- 1.1 days) and in 46 corneas preserved at 4 degrees C for 6.1 +/- 1.9 days in Optisol-GS. The evaluation was repeated after 19.7 +/- 9.1 days of incubation at 31 degrees C in a culture medium containing 2% newborn calf serum.

RESULTS

After the hypothermic storage the corneal endothelium had a mean density of 2475 +/- 159 cells/mm2 without significant difference between the short and the long-term incubation. Several corneas of the two groups showed signs of endothelium degeneration and were positive to trypan blue test. After the incubation at 31 degrees C, the corneas with endothelial degeneration decreased by 52.2% and those positive to trypan blue decreased by 21.7%. Polymorphism (enlarged endothelial cells) increased from 9.6% to 14.5% of the corneas. The remodeling of the endothelium led to a 6.7% decrease in cell density. These results were similar after short-term and long-term storage at 4 degrees C.

CONCLUSIONS

Organ culture was effective in improving corneal endothelium when the hypothermic storage was prolonged to the upper temporal limit for this procedure (7-10 days). These results may encourage the possibility of an eye bank to allocate the available cornea pool, thus decreasing the risk of discarding precious material.

摘要

目的

评估人角膜保存的器官培养方法是否可应用于在4℃下储存数天的角膜。

方法

对140例在4℃下储存至转运至眼库所需最短时间及用于角膜状态初步检查(1.6±1.1天)的人角膜,以及46例在Optisol-GS中于4℃保存6.1±1.9天的角膜,检查角膜内皮细胞的密度、活力和形态。在含2%新生牛血清的培养基中于31℃孵育19.7±9.1天后重复评估。

结果

低温保存后,角膜内皮细胞平均密度为2475±159个/mm²,短期和长期孵育组间无显著差异。两组均有部分角膜出现内皮细胞变性迹象,台盼蓝试验呈阳性。在31℃孵育后,内皮细胞变性的角膜减少了52.2%,台盼蓝阳性的角膜减少了21.7%。多形性(内皮细胞增大)角膜从9.6%增至14.5%。内皮细胞重塑导致细胞密度降低6.7%。4℃短期和长期保存后的结果相似。

结论

当低温保存延长至该方法的时间上限(7 - 10天)时,器官培养对改善角膜内皮有效。这些结果可能促使眼库有机会分配现有的角膜库,从而降低丢弃珍贵材料的风险。

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引用本文的文献

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Commentary: Comparison of structural integrity and functional status of corneal endothelium stored in Cornisol and Optisol-GS.述评:储存在Cornisol和Optisol-GS中的角膜内皮的结构完整性和功能状态比较
Indian J Ophthalmol. 2019 Oct;67(10):1584-1585. doi: 10.4103/ijo.IJO_1107_19.
2
Donor cornea transfer from Optisol GS to organ culture storage: a two-step procedure to increase donor tissue lifespan.从 Optisol GS 向器官培养储存中转移供体角膜:延长供体组织寿命的两步法。
Acta Ophthalmol. 2013 May;91(3):219-25. doi: 10.1111/j.1755-3768.2012.02390.x. Epub 2012 Mar 16.
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The unfolded protein response in human corneal endothelial cells following hypothermic storage: implications of a novel stress pathway.
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Noninvasive optical coherence tomography monitoring of structure and hydration changes of human corneas in different preservation media.不同保存介质下人眼角膜结构和含水量变化的无创光学相干断层扫描监测。
J Biomed Opt. 2011 Feb;16(2):026015. doi: 10.1117/1.3541792.
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Graefes Arch Clin Exp Ophthalmol. 2008 Mar;246(3):369-72. doi: 10.1007/s00417-007-0711-5. Epub 2007 Nov 16.
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