Currently, endothelial keratoplasty is the gold standard for the surgical treatment of Fuchs endothelial corneal dystrophy (FECD). Despite the remarkable success in terms of surgical outcomes, a shortage of corneal donor tissue poses a limitation to performing endothelial keratoplasty in many parts of the world. Cell therapy is a potential alternative strategy to keratoplasty and is currently under investigation. Considering that corneas with FECD may contain relatively healthy endothelial cells, samples obtained by descemetorhexis of eyes undergoing EK for FECD can be used for ex vivo expansion of endothelial cells as an autologous cell culture.

In this study, we established corneal endothelial cell cultures derived from 40 patients that underwent endothelial keratoplasty for advanced FECD. Several parameters were evaluated including patient characteristics such as age, gender, and endothelial cell density as well as various processing and cell culture protocols based on different combinations of shipping temperatures, stabilization periods and treatment methods for corneal endothelial cell dissociation. FECD cultures were classified into three groups as: (i) no cells, (ii) cell cultures with endothelial-like morphology or (iii) cell cultures with fibroblast-like features.

Our data seem to suggest that some factors can influence FECD cell culture characteristics including young age, high paracentral endothelial cell density, low shipping temperature and short stabilization period prior to cell isolation. Treatment with type 1 collagenase for cell isolation can delay endothelial-to-mesenchymal transition, but does not increase proliferative capacity.

Although heterologous corneal endothelial cultures from healthy donors have shown encouraging outcomes, the feasibility of autologous cell therapy as a potential treatment for FECD remains challenging. Low initial cell concentration as well as endothelial to mesenchymal transition are the main obstacles to the application of FECD cultures in the clinical setting.

目前，内皮角膜移植术是 Fuchs 内皮角膜营养不良 (FECD) 手术治疗的金标准。尽管在手术结果方面取得了显着的成功，但角膜供体组织的短缺限制了在世界许多地方进行内皮角膜移植术。细胞疗法是角膜移植术的潜在替代策略，目前正在研究中。考虑到带有 FECD 的角膜可能含有相对健康的内皮细胞，通过对 FECD 进行 EK 的眼睛的脱塞术获得的样本可用于内皮细胞的体外扩增，作为自体细胞培养物。

在这项研究中，我们建立了来自 40 名接受内皮角膜移植术治疗晚期 FECD 的患者的角膜内皮细胞培养物。评估了几个参数，包括患者特征，如年龄、性别和内皮细胞密度，以及基于不同组合的运输温度、稳定期和角膜内皮细胞分离治疗方法的各种处理和细胞培养方案。FECD 培养物分为三组：(i) 无细胞，(ii) 具有内皮样形态的细胞培养物或 (iii) 具有成纤维细胞样特征的细胞培养物。

我们的数据似乎表明，一些因素会影响 FECD 细胞培养特征，包括年轻、中央旁内皮细胞密度高、运输温度低和细胞分离前的稳定期短。用 1 型胶原酶进行细胞分离可以延迟内皮细胞到间充质细胞的转化，但不会增加增殖能力。

尽管来自健康供体的异源角膜内皮培养物已显示出令人鼓舞的结果，但自体细胞疗法作为 FECD 潜在治疗方法的可行性仍然具有挑战性。低初始细胞浓度以及内皮细胞向间充质转化是 FECD 培养物在临床环境中应用的主要障碍。