Transplantation of corneas reconstructed with cultured adult human corneal endothelial cells in nude rats
Transplantation of corneas reconstructed with cultured adult human corneal endothelial cells in nude rats
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DOI：
10.1016/j.exer.2004.05.001
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发表时间：
2004-08-01
影响因子：
3.4
通讯作者：
Yamagami, S
中科院分区：
文献类型：
--
作者：
Mimura, T;Amano, S;Yamagami, S
目的.在裸大鼠模型上检测培养的成人角膜内皮细胞（HCEC）用于角膜重建的可行性。使用无菌棉拭子从刘易斯大鼠的角膜去除内皮细胞。将用荧光标记物氯甲基-苯甲酰氨基二烷基碳菁（CM-Dil）标记的培养的成人HCEC接种到裸露的德斯密氏膜上。将角膜离心，孵育2天后，采用穿透性角膜移植技术移植到裸大鼠眼内（HCEC组）。对照裸角膜接受剥脱内皮且无HCEC的角膜。术后观察28 d，行组织学和荧光显微镜检查。在整个观察期间，HCEC组的平均角膜厚度显著小于对照组。术后28天移植物的角膜内皮细胞密度范围为2425 - 3250个细胞mm（-2）（平均值+/- SD，2744 +/- 337个细胞mm（-2））。术后第28天，角膜内皮素显微镜显示HCEC组角膜后表面有大量DiI标记细胞。冰冻切片显示在后弹力膜上有单层DiI标记细胞。体外培养的人角膜内皮细胞移植于裸鼠后1个月内功能良好，角膜透明性维持良好。(C)2004爱思唯尔有限公司保留所有权利。
Purpose. The feasibility of corneal reconstruction with cultured adult human corneal endothelial cells (HCEC) was examined in a nude rat model.Methods. Endothelial cells were removed from the corneas of Lewis rats using a sterile cotton swab. Cultured adult HCEC labelled with a fluorescent marker chloromethyl-benzamidodialkylcarbocyanine (CM-Dil) were seeded onto the denuded Descemet's membrane. Then the corneas were centrifuged, incubated for 2 days, and transplanted into the eyes of nude rats using the penetrating keratoplasty technique (HCEC group). Control nude received corneas denuded of endothelium and without HCEC. The operated eyes were observed for 28 days after transplantation, and then were subjected to histological and fluorescein microscopic examination.Results. The mean corneal thickness was significantly smaller in the HCEC group than in the control group throughout the observation period. The corneal endothelial cell density of the grafts at 28 days postoperatively ranged from 2425 to 3250 cells mm(-2) (mean +/- SD, 2744 +/- 337 cells mm(-2)). Fluorescein microscopy at 28 days after surgery showed numerous DiI-labelled cells on the posterior corneal surface in the HCEC group. Frozen sections showed a monolayer of DiI-labelled cells on Descemet's membrane.Conclusions. Cultured adult HCEC function well and maintain corneal transparency for 1 month after transplantation in nude rats. (C) 2004 Elsevier Ltd. All rights reserved.