Centre LOEX de l'Université Laval, Génie Tissulaire et Régénération: LOEX-Centre de Recherche FRSQ du Centre Hospitalier affilié Universitaire de Québec, Québec, QC, Canada.
Exp Eye Res. 2012 Jan;94(1):22-31. doi: 10.1016/j.exer.2011.10.018. Epub 2011 Nov 19.
The purpose of this study was to assess the feasibility of initiating primary cultures of corneal endothelial cells from patients suffering from Fuchs endothelial corneal dystrophy (FECD; MIM# 1036800). We also evaluated which conditions yielded the best results for culture. Twenty-nine patients undergoing Descemet stripping automated endothelial keratoplasty consented to the use of their excised Descemet's membrane for this study. Out of the 29 specimens, 18 successfully initiated a culture. Cell morphology varied between endothelial (rounded, slightly elongated cells, n = 12) and fibroblastic-like (thin and very elongated cells, n = 6). These differences in cell morphology were also observed with the normal human corneal endothelial cell cultures. The cultures that initially presented an endothelial morphology maintained their shape in subcultures. Clusterin expression was similar in FECD and normal endothelial cells. Transmission electron microscopy of FECD Descemet's membranes showed a high degree of various abnormalities generally found in this disease, such as a thickened Descemet's membrane, presence of a posterior banded layer, presence of a fibrillar layer and striated bodies of various sizes and periodicities. Patient's age was predictive of culture success, all younger FECD donors generating cultures of endothelial morphology. The absence of a fibrillar layer was also a factor associated with greater success. Culture success was not dependent on specimen size, specimen pigmentation, or patient's preoperative central corneal thickness. In conclusion, this paper shows for the first time that central Descemet's membranes of patients suffering from FECD possess proliferative endothelial cells that can be isolated and cultured without viral transduction, opening the way for new in vitro studies of this disease.
本研究旨在评估从患有 Fuchs 角膜内皮营养不良(FECD;MIM# 1036800)的患者中分离原代角膜内皮细胞的可行性。我们还评估了哪种培养条件能获得最佳结果。29 名接受 Descemet 撕囊自动角膜内皮移植术的患者同意使用他们切除的 Descemet 膜进行本研究。在 29 个标本中,有 18 个成功启动了培养。细胞形态在角膜内皮细胞(圆形,稍长的细胞,n=12)和成纤维细胞样(细长的细胞,n=6)之间有所不同。这种细胞形态的差异也在正常的人角膜内皮细胞培养中观察到。最初表现出内皮形态的培养物在传代培养中保持其形状。FECD 和正常内皮细胞的簇蛋白表达相似。FECD Descemet 膜的透射电子显微镜显示出高度的各种异常,通常在这种疾病中发现,如增厚的 Descemet 膜、后带状层的存在、纤维层的存在以及各种大小和周期性的条纹体。患者年龄是培养成功的预测因素,所有年轻的 FECD 供体均产生具有内皮形态的培养物。无纤维层也是与更高成功率相关的因素。培养成功与否与标本大小、标本色素沉着或患者术前中央角膜厚度无关。总之,本文首次表明,患有 FECD 的患者的中央 Descemet 膜具有增殖性内皮细胞,可以在没有病毒转导的情况下分离和培养,为这种疾病的新的体外研究开辟了道路。