Development of cultivated corneal epithelial cells on amniotic membrane

在严重的眼表面疾病中，例如Stevens-Johnson综合征和眼瘢痕性类天疱疮，其中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、基质瘢痕形成、严重影响视力的新血管形成。在这类疾病中，由于穿透性角膜移植术后的预后差，因此开发了角膜上皮移植的替代手术治疗（角膜缘移植或角膜上皮移植术）以改善眼表重建的结果。最近开发的用于这些疾病的治疗涉及使用培养的角膜上皮干细胞移植来重建由角膜上皮干细胞缺陷引起的眼表面损伤。本课题组建立了以AM为载体的角膜缘上皮细胞培养体系，并进行了动物和人的角膜缘上皮细胞培养。我们还成功地再生角膜上皮细胞在体外使用细胞悬浮培养系统。培养的细胞呈4-5层分层，分化良好，角膜特异性角蛋白3 /12染色，成簇。此后，我们将该系统用于严重眼表疾病的临床应用，并成功实现了眼表重建。在所有病例中，在第一次移植后48小时，角膜表面清晰、光滑，并被移植的同种异体角膜上皮完全覆盖。

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.