Alió Jorge L, Gauthier Benoit R, Gómez Rosado Juan C, Alió Del Barrio Jorge L, Rodríguez Alejandra E, Díaz Corrales Francisco J, Capitán Morales Luis C, Soria Bernat, Lachaud Christian C
Vissum Miranza, Alicante, Spain.
Department of Pathology and Surgery, Area of Ophthalmology, Universidad Miguel Hernandez, Alicante, Spain.
Invest Ophthalmol Vis Sci. 2025 Aug 1;66(11):7. doi: 10.1167/iovs.66.11.7.
To evaluate whether cultured human omental mesothelial cells (OMC) exhibit phenotypic and functional similarities to human corneal endothelial cells (CEC) and whether they can adhere to the corneal stroma and form a biomimetic corneal endothelium when grown on a human anterior lens capsule (HALC).
Human OMC were isolated from the greater omentum. Human B4G12 CEC were used as a reference for native and functional CEC phenotype whereas human mesenchymal stromal cells (MSC) served as a phenotypically distinct control group. OMC, CEC and MSC were compared using flow cytometry, immunofluorescence and qRT-PCR for lineage-specific markers, as well as through a transepithelial permeability assay. The adhesion capacity of OMC was also evaluated on both mouse corneal stroma and a decellularized HALC.
Several cell-surface markers commonly expressed at high levels in MSC (CD13, CD105, CD73 and CD44) were detected at markedly lower levels in OMC and CEC. OMC and CEC, but not MSC, also shared a similar expression profile of key structural markers (pan cytokeratin, ZO-1, β-catenin, N-cadherin, COL4A2, COL8A2) and functional corneal endothelium markers (AQP1, ATP1A1, SLC4A11, CLCN-3). In addition, OMC demonstrated transepithelial permeability values comparable to those of CEC monolayers supporting their barrier-forming capacity. Both CEC and OMC formed cobblestone-like monolayers on mouse corneal stroma and OMC seeded onto HALC generated a bioconstruct that reproduced the key morphological and ultra-structural features of native corneal endothelium.
Overall, our findings show that cultured human OMC exhibit phenotypic and functional similarities to CEC. The ability of OMC to adhere to the corneal stroma and to generate a biomimetic corneal endothelium when combined with HALC highlights their potential use in corneal endothelial regeneration.
评估培养的人网膜间皮细胞（OMC）是否表现出与人类角膜内皮细胞（CEC）相似的表型和功能，以及当它们生长在人晶状体前囊（HALC）上时，是否能够粘附于角膜基质并形成仿生角膜内皮。
从大网膜中分离出人OMC。将人B4G12 CEC用作天然和功能性CEC表型的参考，而人间充质基质细胞（MSC）作为表型不同的对照组。使用流式细胞术、免疫荧光和qRT-PCR对OMC、CEC和MSC进行谱系特异性标志物比较，并通过跨上皮通透性测定进行比较。还在小鼠角膜基质和脱细胞HALC上评估了OMC的粘附能力。
在MSC中通常高水平表达的几种细胞表面标志物（CD13、CD105、CD73和CD44）在OMC和CEC中的表达水平明显较低。OMC和CEC，但不是MSC，也共享关键结构标志物（泛细胞角蛋白、ZO-1、β-连环蛋白、N-钙粘蛋白、COL4A2、COL8A2）和功能性角膜内皮标志物（AQP1、ATP1A1、SLC4A11、CLCN-3）的相似表达谱。此外，OMC的跨上皮通透性值与CEC单层相当，支持其屏障形成能力。CEC和OMC在小鼠角膜基质上均形成鹅卵石样单层，接种到HALC上的OMC产生了一种生物构建体，再现了天然角膜内皮的关键形态和超微结构特征。
总体而言，我们的研究结果表明，培养的人OMC表现出与CEC相似的表型和功能。OMC粘附于角膜基质并与HALC结合时产生仿生角膜内皮的能力突出了它们在角膜内皮再生中的潜在用途。
