Orthotopic corneal transplantation in the mouse - A new surgical technique with minimal endothelial cell loss
Orthotopic corneal transplantation in the mouse - A new surgical technique with minimal endothelial cell loss
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DOI：
10.1007/bf00292359
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发表时间：
1996-11-01
影响因子：
2.7
通讯作者：
Hoffmann, F
中科院分区：
文献类型：
--
作者：
Zhang, EP;Schrunder, S;Hoffmann, F
背景资料：原位进行角膜移植术的小鼠模型是研究同种异体移植反应的重要模型，但小尺寸造成技术上的困难.方法：用共聚焦显微镜和组织学方法测量BALB/c小鼠前夜的前房。10只C3 H小鼠供体角膜分别通过传统技术和新开发的“水下”技术分离，其中打开的供体眼睛在水下不会失去形状。角膜内皮细胞台盼蓝、茜素红S染色。10只BALB/c（H-2d）小鼠接受了用水下技术从C3 H（H-2k）小鼠身上取下的角膜移植片，结果：BALB/c小鼠3.7mm眼角膜直径为3.5mm，角膜中央厚度为170 μ m，上皮占30%，基质占70%。虽然通过新的“水下”技术分离的角膜没有显示出内皮损伤，但使用传统技术发现内皮表面有28 +/- 17.0%的缺陷。所有移植的角膜是明确的，当盖子打开术后2天，第7和第30天（平均16.5天）之间的同种异体移植reaction.Conclusion：新开发的“水下"技术是上级传统的技术，因为浮动的非常薄的供体角膜分离过程中防止内皮缺陷，防止褶皱。通过在小鼠中进行可靠的角膜移植术，该技术促进了对实验性同种异体移植反应的研究。
Background: The murine model of orthotopic performing keratoplasty is important for studying the allograft reaction, but the small dimensions cause technical difficulties.Methods: The anterior chamber of the eve of the BALB/c mouse was measured with the confocal microscope and with histological methods. Ten C3H mouse donor corneas each were separated by the conventional technique and by the newly developed ''underwater'' technique, where the opened donor eye did not lose its shape under water. The corneal endothelium was stained with trypan blue and alizarin red S. Ten BALB/c (H-2d) mice received a corneal graft taken from a C3H (H-2k) mouse by the underwater technique.Results: The 3.7-mm eye of the BALB/c mouse has a corneal diameter of 3.5 mm. The cornea has a central thickness of 170 mu m, the epithelium comprising 30% and the stroma 70%. While none of the corneas separated by the new ''underwater'' technique evidenced endothelial damage, a 28 +/- 17.0% defect of the endothelial surface was found with the conventional technique. All transplanted corneas were clear when the lids were opened on the 2 postoperative day and clouded between the 7th and 30th days (mean 16.5 days) due to an allograft reaction.Conclusion: The newly developed ''underwater'' technique is superior to the conventional technique, since floating of the very thin donor cornea during the separation procedure prevents endothelial defects by guarding against folds. By enabling reliable keratoplasty in the mouse, this technique facilitates studies on the experimental allograft reaction.