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Development of cultivated corneal epithelial cells on amniotic membrane

羊膜上培养角膜上皮细胞的发育

基本信息

批准号：
12557149
负责人：
KINOSHITA Shigeru
金额：
$ 8.32万
依托单位：
Kyoto Prefectural University of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557149/
关键词：
amnion culture corneal epithelium stem cell ocular surface disease ocular surface reconstruction スラムセル

项目摘要

In severe ocular surface diseases, such as Stevens-Johnson syndrome and ocular cicatricial pemphigoid, in which I limbal epithelial cells are destroyed, the neighboring conjunctival epithelial cells invariably cover the corneal surface resulting in chronic inflammation, stromal scarring, neovascularization which severely affects visual acuity. In such diseases, because the prognosis after penetrating keratoplasty is poor, the alternative surgical treatment of corneal epithelial transplantation (limbal transplantation or keratoepithelioplasty) has been developed to improve the outcome of ocular surface reconstruction. The most recently developed treatment for these diseases involves the use of cultivated corneal epithelial stem cell transplantation for reconstructing the ocular surface damage caused by corneal epithelial stem cell deficiencies. Our group developed a corneal limbal epithelial culture system, using AM as a carrier, in an animal and human model. We further have successfully regenerated corneal epithelial cells in vitro using a cell suspension culture system. The cultivated cells showed 4-5 layers of stratification, were well differentiated, and stained with cornea-specific keratin 3 /12 and cluster in. We have since adopted this system for clinical use in severe ocular surface disorders and have successfully achieved ocular surface reconstruction. In all cases, the corneal surfaces were clear, smooth and were perfectly covered with transplanted allo-corneal epithelium at 48 hours after the first transplantation.

在严重的眼表疾病中，如Stevens-Johnson综合征和眼部瘢痕性类天疱疮，角膜缘上皮细胞被破坏，邻近的结膜上皮细胞总是覆盖角膜表面，导致慢性炎症、基质疤痕、新生血管形成，严重影响视力。在此类疾病中，由于穿透性角膜移植术后的预后较差，因此开发了角膜上皮移植（角膜缘移植或角膜上皮成形术）的替代手术治疗，以改善眼表重建的结果。最近开发的针对这些疾病的治疗方法涉及使用培养的角膜上皮干细胞移植来重建因角膜上皮干细胞缺陷引起的眼表损伤。我们的小组在动物和人类模型中开发了一种角膜缘上皮培养系统，使用AM作为载体。我们还利用细胞悬浮培养系统在体外成功地再生了角膜上皮细胞。培养的细胞呈现4-5层分层，分化良好，并用角膜特异性角蛋白3 /12染色并簇入。此后我们将该系统用于临床严重眼表疾病，并成功实现了眼表重建。所有病例的角膜表面均清晰、光滑，并在首次移植后 48 小时被移植的同种角膜上皮完美覆盖。