Establishment of a cultivated human conjunctival epithelium as an alternative tissue source for autologous corneal epithelial transplantation
Establishment of a cultivated human conjunctival epithelium as an alternative tissue source for autologous corneal epithelial transplantation
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DOI：
10.1167/iovs.06-0293
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发表时间：
2006-09-01
影响因子：
4.4
通讯作者：
Kinoshita, Shigeru
中科院分区：
文献类型：
--
作者：
Tanioka, Hidetoshi;Kawasaki, Satoshi;Kinoshita, Shigeru
目的。角膜上皮对于维持角膜透明度至关重要，并且已经努力开发用于角膜缘完全衰竭患者的角膜上皮移植的改进技术。我们评估了移植的培养人结膜上皮 (HCjE) 作为角膜上皮替代物对全角膜和角膜缘缺陷兔的适用性。方法。在人羊膜（AM）上培养至汇合并暴露于气液界面（空气提升）的HCjE细胞被移植到裸露的兔角膜上并监测2周。通过免疫组织化学和透射电子显微镜（TEM）分析培养的HCjE片和移植的上皮。结果。移植的HCjE在随访期间保持透明、光滑且无上皮缺陷。培养的HCjE细胞和移植的上皮均表现出五至六层复层鳞状上皮，其形态与正常角膜上皮相似。基底细胞表达假定的干细胞标记物（ABCG2和P63）以及半桥粒和桥粒组成蛋白。在移植的上皮中发现了细胞角蛋白（CK4、CK13、CK3 和 CK12）和 MUC4。然而，MUC5AC没有表达。结果表明，在 AM 上培养的 HCjE 有潜力用作替代角膜上皮。结论。培养的 HCjE 片的移植是治疗角膜缘衰竭的一种有前途的技术。
PURPOSE. The corneal epithelium is essential for maintaining corneal transparency, and efforts have been made to develop improved techniques for corneal epithelial transplantation in patients with total limbal failure. We evaluated the suitability of transplanted cultivated human conjunctival epithelium ( HCjE) as a corneal epithelium replacement in rabbits with total corneal and limbal deficiency.METHODS. HCjE cells, cultivated on human amniotic membrane ( AM) to confluence and exposed to an air-liquid interface ( air-lifted), were transplanted onto denuded rabbit corneas and monitored for 2 weeks. The cultivated HCjE sheet and the engrafted epithelium were analyzed by immunohistochemistry and transmission electron microscopy ( TEM).RESULTS. The transplanted HCjE remained transparent, smooth, and without epithelial defects during the follow-up period. Both the cultivated HCjE cells and the engrafted epithelium manifested five to six layers of stratified squamous epithelium similar in morphology to normal corneal epithelium. The basal cells expressed the putative stem cell markers ( ABCG2 and P63) and hemidesmosome and desmosome component proteins. The cytokeratins ( CK4, CK13, CK3, and CK12) and MUC4 were found in the engrafted epithelium. However, MUC5AC was not expressed. The results indicate that HCjE cultivated on AM has the potential to be used as an alternative corneal epithelium.CONCLUSIONS. The transplantation of cultivated HCjE sheets is a promising technique for the treatment of eyes with limbal failure.