Phenotypic study of a case with successful transplantation of ex vivo expanded human limbal epithelium for unilateral total limbal stem cell deficiency
Phenotypic study of a case with successful transplantation of ex vivo expanded human limbal epithelium for unilateral total limbal stem cell deficiency
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DOI：
10.1016/s0161-6420(02)01105-3
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发表时间：
2002-08-01
期刊：
影响因子：
13.7
通讯作者：
Tseng, SCG
中科院分区：
文献类型：
--
作者：
Grueterich, M;Espana, EM;Tseng, SCG
目的：为了最大限度地减少单侧角膜缘干细胞缺乏症（LSCD）行结膜角膜缘自体移植术时供体眼的风险，已报道了一种新的方法，即从羊膜（AM）上培养的小角膜缘活检组织中扩增角膜缘上皮祖细胞。在此，我们提出了第一次这样的patient.Design的形态学和表型的结果：干预case report.Methods：一个31岁的男性与严重的酸烧伤，他的左眼接受AM移植在急性期和角膜缘同种异体移植（KLAL）在慢性期的总LSCD。作为治疗失败的KLAL的替代方法，进行了上述新的外科手术。将5.5个月后进行穿透性角膜移植术后获得的角膜纽扣和作为对照的正常角膜纽扣进行苏木精-伊红和免疫荧光染色，以检测角蛋白K3、连接蛋白43、杯状细胞粘蛋白MUC 5AC、层粘连蛋白5和整合素α 3 β 1和α 6 β 4。主要结果测量：临床和免疫组织学特征。所得上皮分层为5 - 6个细胞层，并以与正常角膜上皮相似的方式通过整合素α 3 β 1和α 6 β 4锚定到羊膜基底膜的层粘连蛋白5上。有趣的是，上皮表型是角膜缘，而不是角膜，角蛋白K3和连接蛋白43的基底epitheli.Conclusions的阴性表达的基础上：所描述的技术，确保保存羊膜基底膜，它允许形成的粘附复合物，并保持正常的角膜结构。重建的角膜表面上的角膜缘上皮表型的保存表明，AM提供了一个独特的基质环境，有利于角膜缘上皮祖细胞的保存和扩增。(C)2002年，美国眼科学会。
Objective: To minimize the risk to the donor eye when a conjunctival limbal autograft is performed for unilateral total limbal stem cell deficiency (LSCD), a new approach has been reported of expanding limbal epithelial progenitor cells from a small limbal biopsy cultured on amniotic membrane (AM). Herein, we present for the first time the morphologic and phenotypic outcome of one such patient.Design: Interventional case report.Methods: A 31-year-old male with a severe acid burn to his left eye received AM transplantation at the acute stage and a keratolimbal allograft (KLAL) at the chronic stage for total LSCD. As an alternative to combat the failed KLAL, the above-mentioned new surgical procedure was performed. The corneal button, obtained after a penetrating keratoplasty performed 5.5 months later, and a normal corneal button as a control were submitted to hematoxylin-eosin and immunofluorescence staining for keratin K3, connexin 43, goblet-cell mucin MUC 5AC, laminin 5, and integrins alpha3beta1 and alpha6beta4.Main Outcome Measures: Clinical and immunohistologic features.Results: The resultant epithelium was stratified with five to six cell layers and anchored to laminin 5 of the amniotic basement membrane via integrins alpha3beta1 and alpha6beta4 in a manner similar to the normal corneal epithelium. Intriguingly, the epithelial phenotype was limbal and not corneal, based on the negative expression of keratin K3 and connexin 43 of the basal epithelium.Conclusions: The technique described ensures the preservation of amniotic basement membrane, which allows formation of adhesion complexes and maintains normal corneal architecture. The preservation of a limbal epithelial phenotype on the reconstructed corneal surface indicates that AM provides a unique stromal environment conducive to the preservation and expansion of limbal epithelial progenitor cells. (C) 2002 by the American Academy of Ophthalmology.