Development of cultivated corneal epithelial cells on amniotic membrane

在严重的眼表疾病中，如Stevens-Johnson综合征和眼瘢痕性类天疱疮，其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、间质瘢痕形成、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预后较差，为了改善眼表重建的效果，人们发展了角膜上皮移植的替代手术治疗（角膜缘移植或角膜上皮成形术）。最近发展的治疗这些疾病的方法包括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乏引起的眼表损伤。本小组以AM为载体，在动物和人体模型中建立了角膜缘上皮培养系统。我们进一步成功地利用细胞悬浮培养系统在体外再生角膜上皮细胞。培养的细胞呈4-5层分层，分化良好，具有角膜特异性角蛋白3 /12染色，呈聚集状。此后，我们将该系统应用于严重眼表疾病的临床治疗，并成功实现了眼表重建。在所有病例中，第一次移植后48小时，角膜表面清晰、光滑，并被移植的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.