Di Girolamo N, Tedla N, Kumar R K, McCluskey P, Lloyd A, Coroneo M T, Wakefield D
Inflammation Research Unit, School of Pathology, University of New South Wales, Australia.
Br J Ophthalmol. 1999 Sep;83(9):1077-82. doi: 10.1136/bjo.83.9.1077.
BACKGROUND/AIMS: Pterygia are a common disorder of the ocular surface. The disease represents a chronic fibrovascular and degenerative process thought to originate at the conjunctival-corneal junction, where altered limbal stem cells are proposed to be the cell of origin. Extensive epidemiological evidence exists to implicate ultraviolet B irradiation in the pathogenesis of pterygia. To date no animal or in vitro culture model has been developed to test such an hypothesis. The aim of this study was to establish and characterise a pure population of epithelial cells derived from pterygium tissue.
Tissue specimens were obtained from patients undergoing pterygium excision. Explants were cultured in either serum free or serum supplemented medium. Primary and passaged cells were processed for light microscopy, analysed by flow cytometry, and characterised immunohistochemically using specific antibodies.
In serum free culture, cuboidal cells with typical morphology of epithelial cells migrated from the pterygium explants from 3 days onwards and eventually formed a cohesive monolayer. Passaged cells consisted of 98.4% cytokeratin positive cells and demonstrated immunoreactivity for multiple cytokeratins, including AE1, AE3, AE5, but were negative for AE8. These cells also expressed an epithelial specific antigen, together with vimentin and mucin, as did epithelial cells in sections of pterygia.
A relatively simple method of isolating pterygium epithelial cells has been established. Cultured pterygium epithelial cells are phenotypically and functionally similar to their in vivo counterparts with respect to keratin, vimentin, and mucin expression. In vitro assays using these cells may aid in elucidating the pathogenesis of pterygia.
背景/目的：翼状胬肉是一种常见的眼表疾病。该疾病是一种慢性纤维血管性和退行性病变，被认为起源于结膜角膜交界处，此处改变的角膜缘干细胞被认为是起源细胞。有大量流行病学证据表明紫外线B照射与翼状胬肉的发病机制有关。迄今为止，尚未建立动物或体外培养模型来验证这一假说。本研究的目的是建立并鉴定源自翼状胬肉组织的纯上皮细胞群体。
从接受翼状胬肉切除术的患者获取组织标本。外植体在无血清或补充血清的培养基中培养。对原代细胞和传代细胞进行光学显微镜检查、流式细胞术分析，并使用特异性抗体进行免疫组织化学鉴定。
在无血清培养中，具有典型上皮细胞形态的立方体细胞从第3天起从翼状胬肉外植体迁移出来，最终形成紧密的单层。传代细胞中98.4%为细胞角蛋白阳性细胞，对多种细胞角蛋白包括AE1、AE3、AE5呈免疫反应性，但对AE8呈阴性。这些细胞还表达上皮特异性抗原，以及波形蛋白和黏蛋白，翼状胬肉切片中的上皮细胞也是如此。
已建立了一种相对简单的分离翼状胬肉上皮细胞的方法。培养的翼状胬肉上皮细胞在角蛋白、波形蛋白和黏蛋白表达方面在表型和功能上与其体内对应细胞相似。使用这些细胞进行的体外试验可能有助于阐明翼状胬肉的发病机制。
