Prospects for Descemet Stripping Automated Endothelial Keratoplasty Using Cultured Human Corneal Endothelial Cells
Prospects for Descemet Stripping Automated Endothelial Keratoplasty Using Cultured Human Corneal Endothelial Cells
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DOI：
10.4172/2161-0991.s2-001
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发表时间：
2013
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影响因子：
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通讯作者：
T. Mimura;S. Yamagami;Seiichi Yokoo;T. Usui;S. Amano
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作者：
T. Mimura;S. Yamagami;Seiichi Yokoo;T. Usui;S. Amano
研究背景：后弹力层剥脱式自动角膜内皮移植术（DSAEK）允许选择性置换病变角膜内皮。然而，DSAEK需要供体角膜，并且全球角膜短缺限制了其应用。本文介绍了我们近年来在利用培养的人角膜内皮细胞（HCEC）进行组织工程化DSAEK方面的工作以及最近发表的关于HCEC前体的实验数据。方法和结果：我们将DiI标记的培养HCEC接种到胶原片上，得到HCEC片。这些片的泵功能参数是人供体角膜的泵功能参数的76%至95%。新西兰白色兔股后弹力层断裂后，将HCEC片移植于兔股后基质上（DSAEK组）。仅使用后弹力层破裂的兔角膜为对照组。在整个观察期间，DSAEK组的平均角膜厚度显著小于未移植对照组。DSAEK组角膜后表面有DiI标记细胞。苏木精-伊红染色显微镜下观察到对照组有严重的间质水肿，而DSAEK组无。接下来，我们从人供体角膜中分离出HCEC前体。培养的前体细胞形成表达神经和间充质蛋白的球形集落。这些集落的后代是HCEC样六边形细胞。使用HCEC前体构建的细胞片显示出比用分化培养的HCEC构建的细胞片更强的BrdU和巢蛋白染色。结论：这些发现表明，从成人供体角膜移植的培养的HCECs保留了其角膜脱水功能，并表明用HCECs进行DSAEK治疗内皮功能障碍的可行性。成人角膜内皮细胞含有可分化为角膜内皮细胞的前体细胞。HCEC前体可能成为构建HCEC包被胶原片的有力工具。* 通讯作者：Tatsuya Mimura，Department of Ophthalmology，University of Tokyo Graduate School of Medicine，7-3-1 Hongo，文京区，Tokyo，113-8655 Japan，电话：+81 35800-5108（Ex 33503）;传真：+81 33817-0798;电子邮件：mimurat-tky@umin.ac.jp接收日期：2011年9月24日;接受日期：2011年11月11日;发表于2011年11月15日引文：Mimura T，Yamagami S，Yokoo S，Amano S，Amani T（2011）使用培养的人角膜内皮细胞进行后弹力层剥离自动内皮角膜移植术的前景。J Transplant Technol Res S2：001.版权所有：上海市徐汇区宜山路108号© 2011 Mimura T，et al.这是一篇开放获取的文章，根据知识共享署名许可证的条款分发，该许可证允许在任何媒体上无限制地使用，分发和复制，前提是原作者和来源被记入贷方。
Study background: Descemet stripping automated endothelial keratoplasty (DSAEK) allows selective replacement of the diseased corneal endothelium. However, DSAEK requires a donor cornea and the worldwide shortage of corneas limits its application. In this review, we introduce our recent work on tissue engineering for DSAEK using cultured human corneal endothelial cells (HCEC) and recently published experimental data on HCEC precursors. Methods and Results: We seeded DiI-labeled cultured HCECs onto collagen sheets, yielding HCEC sheets. The pump function parameters of these sheets were 76% to 95% of those for human donor corneas. Then HCEC sheets were transplanted onto the posterior stroma of New Zealand white rabbits after Descemetorhexis (DSAEK group). Rabbit corneas with only Descemetorhexis were the control group. The mean corneal thickness was significantly smaller in the DSAEK group than in the untransplanted control group throughout the observation period. DiI-labeled cells covered the posterior corneal surface in the DSAEK group. Severe stromal edema was detected in the control group by microscopy with hematoxylin-eosin staining, but not in the DSAEK group. Next, we isolated HCEC precursors from human donor corneas. Cultured precursor cells formed sphere colonies that expressed neural and mesenchymal proteins. The progeny of these colonies were HCEC-like hexagonal cells. Cell sheets constructed using HCEC precursors showed stronger staining for BrdU and nestin than cell sheets constructed with differentiated cultured HCECs. Conclusions: These findings indicate that cultured HCECs transplanted from adult human donor corneas retain their corneal dehydration function and suggest the feasibility of performing DSAEK with HCECs to treat endothelial dysfunction. Adult human corneal endothelium contains precursors that can differentiate into corneal endothelial cells. HCEC precursors may become a powerful tool for the construction of HCEC-coated collagen sheets. *Corresponding author: Tatsuya Mimura, Department of Ophthalmology, University of Tokyo Graduate School of Medicine, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-8655 Japan, Tel: +81 35800-5108 (Ex33503); Fax: +81 33817-0798; E-mail: mimurat-tky@umin.ac.jp Received September 24, 2011; Accepted November 11, 2011; Published November 15, 2011 Citation: Mimura T, Yamagami S, Yokoo S, Usui T, Amano S (2011) Prospects for Descemet Stripping Automated Endothelial Keratoplasty Using Cultured Human Corneal Endothelial Cells. J Transplant Technol Res S2:001. doi:10.4172/21610991.S2-001 Copyright: © 2011 Mimura T, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.