Surgical Rehabilitation of Ocular Surface : Epithelial Transplantation

眼表手术康复：上皮移植

基本信息

批准号：
04454442
负责人：
KINOSHITA Shigeru
金额：
$ 2.05万
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 General Scientific Research (B)
财政年份：
1992
资助国家：
日本
起止时间：
1992 至 1993
项目状态：
已结题

项目摘要

We aimed at developing effective treatment for primary ocular surface disorders. For this, in this project, we mainly focused on the methods controlling proliferation and differentiation of corneal epithelial cells. First, we established a noble cell culture system for freeze and long-term subculture of rabbit corneal epithelial cells. This culture media is composed of very low Ca^<2+> concentration with high aminoacid contents. Second, we established an immortalized rabbit corneal epithelial cell line. For this, primary cultured rabbit corneal epithelial cells were infected with a recombinant SV40-adenovirus vector and were cloned. This cell line shares properties consistent with normal corneal epithelial cells. Although these cells were not transferable as a epithelial transplantation at this point, our success has made a marked progress in handling corneal epithelial cells in vitro. Third, we investigated biological roles of several cell growth and/or regulatory factors. For instance, epidermal growth factor, which mainly comes from lacrimal gland, promoted corneal epithelial cell growth. Similar phenomenon was found in keratinocyte growth factor, but this was produced only in the corneal stromal cells. On the other hand, transforming growth factor -beta did inhibit corneal epithelial cell growth. We also studied spartial distribution of three TGF-beta isoforms in the cornea, and found TGF- beta 2 predominantly.In the study of epithelial allograft rejection in vivo, we are now investigating IL-2 levels in tears in patients with cyclosporin treatment after corneal epithelial transplantation.

我们旨在开发针对原发性眼表面疾病的有效治疗方法。为此，在这个项目中，我们主要关注控制角膜上皮细胞增殖和分化的方法。首先，我们建立了一个高贵的细胞培养系统，用于冻结和长期的兔角膜上皮细胞的长期亚培养。该培养基由非常低的Ca^<2+>浓度组成，具有较高的氨基酸含量。其次，我们建立了永生的兔角膜上皮细胞系。为此，用重组SV40-腺病毒载体感染原代兔角膜上皮细胞，并克隆。该细胞系具有与正常角膜上皮细胞一致的特性。尽管这些细胞在这一点上不能作为上皮移植转移，但我们的成功在体外处理角膜上皮细胞方面取得了显着的进展。第三，我们研究了几种细胞生长和/或调节因素的生物学作用。例如，主要来自泪腺的表皮生长因子促进了角膜上皮细胞的生长。在角质形成细胞生长因子中发现了类似的现象，但这仅在角膜基质细胞中产生。另一方面，转化生长因子-beta确实抑制了角膜上皮细胞的生长。我们还研究了角膜中三种TGF-β同工型的斯巴达分布，并主要发现了TGF-beta 2。在体内的上皮同种异体移植排斥反应的研究中，我们现在正在研究角膜上皮上皮移植后环孢子治疗患者的泪液中的IL-2水平。